Patent application title: NOVEL BACTERIOPHAGE AND ANTIBACTERIAL COMPOSITION COMPRISING THE SAME
Inventors:
Hyo Seel Seo (Wonju-Si, KR)
Eun Mi Shin (Seoul, KR)
Gi Duk Bae (Seoul, KR)
Jae-Won Kim (Yongin-Si, KR)
Jae-Won Kim (Yongin-Si, KR)
Assignees:
CJ CHEILJEDANG CORPORATION
IPC8 Class: AC12N700FI
USPC Class:
424 936
Class name: Drug, bio-affecting and body treating compositions whole live micro-organism, cell, or virus containing virus or bacteriophage
Publication date: 2016-03-24
Patent application number: 20160083695
Abstract:
Provided is a novel bacteriophage ΦCJ23 (KCCM11365P). In addition,
the present invention relates to an antibacterial composition including
the bacteriophage ΦCJ23 (KCCM11365P) as an active ingredient.
Further, provided is a method of preventing and/or treating infectious
diseases by avian pathogenic Escherichia coli (APEC) in birds using the
bacteriophage ΦCJ23(KCCM11365P) or the antibacterial composition
containing the bacteriophage ΦCJ23(KCCM11365P) as an active
ingredient.Claims:
1. A novel bacteriophage ΦCJ23 (KCCM11365P) having a specific
bactericidal activity against Avian Pathogenic Escherichia coli (APEC).
2. A composition for preventing or treating an infectious disease caused by APEC, comprising the bacteriophage ΦCJ23 (KCCM11365P) of claim 1 as an active ingredient.
3. The composition according to claim 2, wherein the infectious disease is avian colibacillosis.
4. An antibiotic comprising the bacteriophage ΦCJ23 (KCCM11365P) of claim 1 as an active ingredient.
5. A feed additive or drinking water additive for birds comprising the bacteriophage ΦCJ23 (KCCM11365P) of claim 1 as an active ingredient.
6. A disinfectant or cleaner comprising the bacteriophage ΦCJ23 (KCCM11365P) of claim 1 as an active ingredient.
7. A method of preventing or treating an infectious disease caused by APEC, comprising administering the bacteriophage ΦCJ23 (KCCM11365P) of claim 1 to birds.
8. The method according to claim 7, wherein the infectious disease is avian colibacillosis.
9. A method of preventing or treating an infectious disease caused by APEC, comprising administering the composition of claim 2 to birds.
10. The method according to claim 9, wherein the infectious disease is avian colibacillosis.
Description:
TECHNICAL FIELD
[0001] The present invention relates to a novel bacteriophage having a specific bacteriocidal activity against avian pathogenic Escherichia coli (APEC) and antibacterial composition comprising the same. In addition, the present invention relates to a method of preventing or treating poultry diseases using the novel bacteriophage or the antibacterial composition.
BACKGROUND ART
[0002] Escherichia coli (hereinafter referred to as `E. coli`) is a Gram-negative, short rod-shaped bacterium belonging to the genus Escherichia and the family Enterobacteriaceae, and is one of the normal flora existing in the intestines of various animals including mammals. It was known that most of the strains of E. coli are non-pathogenic and may cause opportunistic infections, but some highly pathogenic strains cause diverse intestinal diseases and septicemia in animals including humans.
[0003] It was known that among these E. coli strains, particularly, avian pathogenic E. coli (APEC), which is E. coli infected through respiratory tract of birds, for example, chickens, ducks, turkeys, or the like, infiltrates into the body through respiratory mucosa. APEC causes various diseases such as septicemia, granuloma, airsacculitis, salpingitis, arthritis, or the like, in birds. Particularly, APEC causes significant economic damage to a poultry industry in that APEC causes respiratory diseases mainly in poultry, or the like, such that APEC becomes a problem.
[0004] Meanwhile, bacteriophage is a specialized type of virus that infects and destroys only bacteria, and can self-replicate only inside host bacteria. The bacteriophage has strong host specificity as compared to antibiotics, and recently, a problem of emergence of resistant bacteria against use of antibiotics has been serious, such that an interest in practical use of the bacteriophage has increased (Non-Patent Documents 1 and 2).
[0005] Therefore, research regarding the bacteriophage has been actively conducted in many countries around the world, and in addition to a patent application for bacteriophage, an attempt to acquire Food and Drug Administration (FDA) approval for a composition containing the bacteriophage has been gradually increased.
[0006] As the prior art for the bacteriophage, 7 kinds of bacteriophages for controlling E. coli 0157:H have been disclosed in Patent Document 1, and a bacteriophage having a specific bacteriocidal activity against Staphylococcus aureus has been disclosed in Patent Document 2. Further, lytic protein derived from a bacteriophage specifically destroying a peptidoglycan structure of bacterial cell membrane, and bacteria lysates by the lytic protein have been disclosed in Patent Document 3.
[0007] However, in spite of presence of the following prior arts, a technology associated with the bacteriophage for preventing and/or treating infectious diseases by APEC, which is an important problem in breeding birds including poultry, is still insufficient, such that a bacteriophage and a technology associated with the bacteriophage should be developed.
PRIOR ART DOCUMENT
Patent Document
[0008] (Patent Document 1) U.S. Pat. No. 6,485,902
[0009] (Patent Document 2) Korea Patent Registration No. 10-0910961 B1
[0010] (Patent Document 3) Korean Patent Laid-Open Publication No. 10-2009-0021475 A
Non-Patent Document
[0011] (Non Patent Document 1) Cislo M, et al., Arch. Immunol. Ther. Exp. 2:175-183, 1987
[0012] (Non Patent Document 2) Sung Hun Kim et al, Bacteriophage, Novel Alternative Antibiotics, BioWave Vol. 7 No. 15, 2005, BRIC
DISCLOSURE
Technical Problem
[0013] The present inventors conducted studies in order to solve problems such as resistant bacteria occurring upon the use of antibiotics, and antibiotics remaining in meat, and the like, and efficiently prevent and treat infectious diseases in birds. As a result, the present inventors isolated new bacteriophage ΦCJ23 (KCCM11365P) having a specific bacteriocidal activity against APEC, causing respiratory diseases in poultry, from nature.
[0014] In addition, the present inventors identified morphological, biochemical, and genetic characteristics of the novel bacteriophage, and confirmed that the bacteriophage had excellent acid resistance, heat resistance, and the like, thereby developed an antibiotic, a disinfectant, a feed additive, and other compositions using the novel bacteriophage. Further, the present inventors developed a composition for preventing or treating infectious diseases generating in birds, and a method of preventing or treating the disease using the composition.
[0015] The present invention provides a novel bacteriophage ΦCJ23 (KCCM11365P) having a specific bactericidal activity against APEC.
[0016] In addition, the present invention provides a composition of preventing and/or treating infectious diseases by APEC containing the bacteriophage ΦCJ23 (KCCM11365P) as an active ingredient.
[0017] Further, the present invention provides an antibiotic, a feed additive, a drinking water additive, a disinfectant, or a cleaner containing the bacteriophage ΦCJ23 (KCCM11365P) as an active ingredient.
[0018] Furthermore, the present invention provides a method of preventing and/or treating infectious diseases by APEC using the bacteriophage ΦCJ23 (KCCM11365P) or a composition containing the bacteriophage ΦCJ23 (KCCM11365P) as an active ingredient.
Technical Solution
[0019] An exemplary embodiment of the present invention provides a novel bacteriophage ΦCJ23 (KCCM11365P) having a specific bactericidal activity against avian pathogenic Escherichia coli (APEC).
[0020] Another exemplary embodiment of the present invention provides a composition for preventing or treating an infectious disease caused by APEC, containing the bacteriophage ΦCJ23 (KCCM11365P) as described above as an active ingredient.
[0021] Another exemplary embodiment of the present invention provides an antibiotic, a feed additive, a drinking water additive, a disinfectant, or a cleaner containing the bacteriophage ΦCJ23 (KCCM11365P) as described above as an active ingredient.
[0022] Another exemplary embodiment of the present invention provides a method of preventing or treating an infectious disease caused by APEC, the method including administering the bacteriophage ΦCJ23 (KCCM11365P) or the composition containing the bacteriophage ΦCJ23 as described above as an active ingredient to birds.
Advantageous Effects
[0023] The bacteriophage ΦCJ23 (KCCM11365P) according to the present invention has the specific bactericidal activity against avian pathogenic Escherichia coli (APEC).
[0024] In addition, since the bacteriophage ΦCJ23 (KCCM11365P) of the present invention has excellent acid resistance, heat resistance, and drought resistance, it may not only be used as a material for preventing or treating infectious diseases by APEC in various temperature or pH ranges, but also utilized as an antibiotic, a feed additive, a drinking water additive, a disinfectant, and a cleaner, or the like.
[0025] Further, according to the present invention, infectious diseases by APEC may be prevented or treated by administering the bacteriophage ΦCJ23 (KCCM11365P) or a composition containing the bacteriophage ΦCJ23 (KCCM11365P) as an active ingredient to birds.
DESCRIPTION OF DRAWINGS
[0026] FIG. 1 is an electron microscope photograph of a novel bacteriophage ΦCJ23 (KCCM11365P, hereinafter, referred to as `ΦCJ23`).
[0027] FIG. 2 shows a pulsed field gel electrophoresis (PFGE) result of the novel bacteriophage ΦCJ23.
[0028] FIG. 3 shows a sodiumdodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) result of the novel bacteriophage ΦCJ23.
[0029] FIG. 4 is a graph showing a result of an acid resistance test of the novel bacteriophage ΦCJ23.
[0030] FIG. 5 is a graph showing a result of a heat resistance test of the novel bacteriophage ΦCJ23.
BEST MODE
[0031] Hereinafter, the present invention will be described in detail. Since contents that are not described in the present specification may be sufficiently recognized and inferred by those skilled in the art or similar art, a description thereof will be omitted.
[0032] In detail, one embodiment of the present invention provides a novel bacteriophage ΦCJ23 (KCCM11365P) having a specific bacteriocidal activity against avian pathogenic Escherichia coli (APEC).
[0033] APEC, which is E. coli infected through respiratory tract of birds such as, chickens, ducks, turkeys, or the like, infiltrates into bodies of birds through respiratory mucosa to cause various diseases such as septicemia, granuloma, air sacculitis, salpingitis, arthritis, or the like. APEC is a Gram-negative, rod-shaped bacterium similarly to a general E. coli, and has motility due to peritrichous flagella, and is an aerobic or facultative anaerobic bacterium decomposing lactose or fructose to produce acid and gas.
[0034] APEC well grows in a general medium, and may grow at about 7 to 48° C., and an optimal growth temperature is about 35 to 37° C. Particularly, virulence factors are effectively expressed at about 42° C., which is close to a body temperature of the bird. In addition, APEC may grow in a pH range of 4.5 to 9.0.
[0035] A bacteriophage is a bacteria-specific virus infecting specific bacteria to suppress and inhibit growth of the bacteria, and means a virus including single or double stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) as a genetic material.
[0036] The bacteriophage ΦCJ23 of the present invention, which is a species-selective bacteriophage that selectively infects APEC, has a structure of an isometric capsid but a tail is not observed (FIG. 1), and morphologically belongs to Podoviridae.
[0037] The bacteriophage ΦCJ23, which was a bacteriophage newly isolated by the present inventors, was deposited at Korean Culture Center of Microorganisms (361-221, Hongjedong, Seodamun-gu, Seoul, Korea) as a deposition number KCCM11365P on Jan. 30, 2013.
[0038] In another embodiment, the present invention provides a composition for prevention or treating infectious diseases by APEC containing the bacteriophage ΦCJ23 as an active ingredient.
[0039] Since the bacteriophage ΦCJ23 has an antibacterial activity capable of specifically killing APEC, it may be used to prevent or treat diseases generated by infection of APEC. As a preferable example of the infectious diseases by APEC, there is avian colibacillosis, but the present invention is not limited thereto.
[0040] The avian colibacillosis, which is a disease generated when the respiratory track of birds, or the like, is infected by pathogenic E. coli, causes various lesions such as airsacculitis, perihepatitis, peritonitis, pericarditis, salpingitis, omphalitis, osteomyelitis, or septicemia, or the like, thereby inhibits growth and causes mortality of the infected birds.
[0041] The term "prevention" as used herein, refers to all actions of providing the bacteriophage ΦCJ23 and/or the composition comprising the bacteriophage ΦCJ23 as the active ingredient to targets, to suppress the corresponding disease or retard disease occurring.
[0042] The term "treatment" as used herein, refers to all actions of providing the bacteriophage ΦCJ23 and/or the composition comprising the bacteriophage ΦCJ23 as the active ingredient to targets, to thereby allow the symptom of the corresponding disease caused by infection to get better or to be alleviated.
[0043] The composition for preventing or treating the infectious disease caused by APEC according to the present invention may contain the bacteriophage ΦCJ23 in an amount of preferably 5×102 to 5×1012 pfu/ml, more preferably, 1×106 to 1×1010 pfu/ml.
[0044] The composition for preventing or treating the infectious disease caused by APEC according to the present invention may further contain a pharmaceutically acceptable carrier, and be formulated together with the carrier to be provided as food, a drug, a feed additive, or a drinking water additive, etc. The term "pharmaceutically acceptable carrier" as used herein, means a carrier or a diluent that does not stimulate living organism nor inhibit biological activity and properties of an administered compound.
[0045] A kind of carrier usable in the present invention is not particularly limited, and any carrier may be used as long as it is generally used in the art and is pharmaceutically acceptable. As a non-restrictive example of the carrier, there are saline, sterile water, Ringer's solution, buffered saline, an albumin injection solution, a dextrose solution, a maltodextrin solution, glycerol, ethanol, and the like. These may be used alone or as a mixture of at least two of these.
[0046] In addition, if necessary, another general additive such as an antioxidant, a buffer, and/or a bacteriostatic agent, etc., may be further added and be used, and the composition may be formulated into a formulation for injection such as an aqueous solution, suspension, emulsion, or the like, pills, capsules, granules, tablets, or the like by additionally adding a diluent, a dispersant, a surfactant, a binder, and/or a lubricant, etc., and then be used.
[0047] An administration method of the composition for preventing or treating infectious diseases by APEC of the invention is not particularly limited, but any method generally used in the art may be used. As a non-restrictive example of the administration method, the composition may be orally or parenterally administered.
[0048] As a non-restrictive example of the formulation for oral administration, there are troches, lozenge, tablets, aqueous suspensions, oily suspensions, prepared powder, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs, etc.
[0049] In order to formulate the composition according to the present invention into a formulation such as a tablet, or a capsule, etc., the formulation may further contain a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose, or gelatin; an excipient such as dicalcium phosphate, or the like; a disintegrant such as corn starch, or sweet potato starch, or the like; a lubricant such as magnesium stearate, calcium stearate, sodium stearyl fumarate, polyethylene glycol wax, or the like. In the case of the capsule formulation, the formulation may further contain a liquid carrier such as fatty oil in addition to the above-mentioned materials.
[0050] As a parenteral administration method of the invention, an intravenous administration method, an abdominal administration method, an intramuscular administration method, a subcutaneous administration method, or a local administration method, etc., may be used. In addition, a method of applying or spraying the composition onto a disease site may also be used, but the present invention is not limited thereto.
[0051] An example of the formulation for the parenteral administration may include formulations for injection such as subcutaneous injection, intravenous injection, intramuscular injection, or the like; suppository formulations; or spray formulations such as aerosol formulations capable of being inhaled through respiratory track, or the like, but the present invention is not limited thereto. In order to formulate the composition into the injection formulation, the composition according to the present invention may be mixed with a stabilizer or a buffer in water to thereby prepare a solution or suspension, and then, the prepared solution or suspension may be formulated in a unit dose for an ampoule or vial. In the case of formulating the composition into the spray formulation such as the aerosol formulation, or the like, a propellant, or the like, may be mixed together with an additive so that a water-dispersed condensate or wet powder could be dispersed.
[0052] A suitable application, spray, or administration dose of the composition in the invention for preventing or treating infectious diseases by APEC may be variously determined depending on factors such as age, weight, sex, degree of symptom of disease, ingesting food, excretion rate of administration subject animals, as well as a method of formulating the composition, an administration method, an administration time and/or route. Generally, a veterinarian having ordinary skill in the art may easily determine and prescribe an effective dose for the desired treatment.
[0053] In another embodiment, the present invention provides an antibiotic comprising the bacteriophage ΦCJ23 as an active ingredient.
[0054] The term "antibiotic" as used herein, means an agent capable of being provided to subjects including humans in a drug form to thereby kill bacteria, and corresponds to a concept collectively indicating a preservative, a disinfectant, and an antibacterial agent.
[0055] The antibiotic containing the bacteriophage ΦCJ23 according to the present invention as the active ingredient, may advantage having high specificity to APEC as compared to conventional antibiotic, thereby not killing beneficial bacteria but killing specific pathogenic bacterial, and does not induce drug resistance, so that the antibiotic according to the present invention may be provided as a novel antibiotic having an elongated lifespan as compared to conventional antibiotic.
[0056] In another embodiment, the present invention provides a feed additive and a drinking water additive for birds, particularly, poultry containing the bacteriophage ΦCJ23 (KCCM11365P) as an active ingredient.
[0057] The term "poultry" as used herein, is a concept collectively indicating animals belonging to birds among domestic animals. The poultry is not particularly limited, but may include, preferably, at least one selected from a groups consisting of chickens, ducks, and turkeys.
[0058] The feed additive or the drinking water additive for birds according to the present invention may be used in a manner in which the bacteriophage ΦCJ23 or the composition containing the smae is individually prepared in a feed additive or drinking water additive form, and then mixed with a feed or drinking water, or in a manner in which the bacteriophage ΦCJ23 or the composition containing the smae my be directly added at the time of preparing the feed or the drinking water.
[0059] The bacteriophage ΦCJ23 or the composition containing the smae used as the feed additive or drinking water additive according to the present invention, may be in a liquid state or dried state, and preferably, in a dried powder form.
[0060] A drying method for preparing the feed additive and the drinking water additive according to the present invention in the dried powder form is not particularly limited, but a method generally used in the art may be used. As a non-restrictive example of the drying method, there are an air drying method, natural drying method, a spray drying method, a freeze-drying method, or the like. These methods may be used alone or at least two methods may be used together with each other.
[0061] Another non-pathogenic microbe may be additionally added to the feed additive or drinking water additive of the invention. A non-restrictive example of the microbe capable of being added may be selected from a group consisting of bacillus subtilis, capable of producing protease, lipase, and/or sugar converting enzyme such as bacillus subtilis, or the like; Lactobacillus sp. having physiological activity and degradation activity for an organic material under anaerobic conditions such as cow's stomach; mold fungi having effects of increasing weight of domestic animal, milk yield, and digestibility of the feed such as Aspergillus oryzae, or the like; and yeasts such as Saccharomyce scerevisiae, or the like. These may be used alone or as a mixture of at least two of these.
[0062] The feed additive or the drinking water additive containing the bacteriophage ΦCJ23 according to the present invention as the active ingredient may further contain other additives, as needed. As a non-restrictive example of the usable additive, there are a binder, an emulsifier, a preservative, and the like, which are added in order to prevent quality of the feed or driving water from being deteriorated; amino acids, vitamins, enzymes, probiotics, flavoring agents, non-protein nitrogen compounds, silicates, buffers, coloring agents, extractants, or oligosaccharides, and the like, which are added in order to increase utility of the feed or drinking water. Otherwise, the additive may further include a feed mixing agent, or the like. These may be used alone or as a mixture of at least two of these.
[0063] The feed additive may be contained at a content of 0.05 to 10, more preferably 0.1 to 2 parts by weight based on 100 parts by weight of the feed. The drinking water additive of the invention may be contained at a content of 0.0001 to 0.01, more preferably 0.001 to 0.005 parts by weight based on 100 parts by weight of the drinking water. The activity of the bacteriophage ΦCJ23 against APEC may be sufficiently exhibited in the above-mentioned range.
[0064] In another embodiment, the present invention provides a feed or drinking water prepared by adding a feed additive or a drinking water additive containing the bacteriophage ΦCJ23 as an active ingredient or directly adding the bacteriophage ΦCJ23.
[0065] The feed used in the present invention is not particularly limited, but any feed generally used in the art may be used. A non-restrictive example of the feed may include plant feeds such as grains, roots and fruit, food processing byproducts, algaes, fiber, pharmaceutical byproducts, fats, starches, cucurbitaceous, or grain byproducts; and animal feeds such as proteins, inorganic materials, fats, minerals, single cell proteins, animal planktons, or foods. These may be used alone or as a mixture of at least two of these.
[0066] The drinking water used in the present invention is not particularly limited, but any drinking water generally used in the present invention may be used.
[0067] In another embodiment, the present invention may provide a disinfectant or a cleaner containing the bacteriophage ΦCJ23 as an active ingredient. A formulation of the disinfectant or cleaner is not particularly limited, it may be formulated into any formulation known in the art.
[0068] The disinfectant may be sprayed in order to remove APEC, and can be sprayed onto a region in which birds live, a slaughterhouse, a mortality area, a cooking place or cooking equipment, or the like, but the present invention is not limited thereto.
[0069] The cleaner may be used to wash surface of the skin or each of the sites of bodies of birds exposed or to be exposed to APEC, but the present invention is not limited thereto.
[0070] In another embodiment, the present invention provides a method of preventing or treating infectious diseases by APEC by using the bacteriophage ΦCJ23 or the composition comprising the same as an active ingredient.
[0071] In detail, the method of preventing or treating infectious diseases according to the present invention, may include administering the bacteriophage ΦCJ23 or the composition containing the smae as the active ingredient to birds infected by APEC or being at risk of infection of APEC in a pharmaceutically effective dose. It will be apparent to those skilled in the art that when the pharmaceutical composition is administered to patient, the suitable total daily dose may be determined by an attending physician or veterinarian within the scope of sound medical judgement.
[0072] A specific pharmaceutically effective dose of the bacteriophage ΦCJ23 or the composition containing the smae as the active ingredient for a specific bird, may be determined considering an administration time and an administration route of the bacteriophage ΦCJ23 or the composition containing the same, a secretion rate of the composition, a therapy duration period, or the like, in addition to a kind and a degree of the desired response, age, weight, general healthy state, sex, or diet of the corresponding individual. In addition, the pharmaceutically effective dose may be variously changed according to various factors such as ingredients of drugs or other compositions simultaneously or separately used and similar factors well known in a medical field.
[0073] The bacteriophage ΦCJ23 according to the present invention or the composition containing the same as the active ingredient may be administered as a pharmaceutical form (nasal spray) to birds or administered in a method of directly added to a feed or drinking water of the birds and then feeding the feed or drinking water. In addition, the bacteriophage ΦCJ23 or the composition containing the same may be mixed in a feed or drinking water in a form of a feed additive or drinking water additive and then administered.
[0074] The administration route and administration method of the bacteriophage ΦCJ23 according to the present invention or the composition containing the same as the active ingredient are not particularly limited, but any administration route and administration method may be used as long as the bacteriophage ΦCJ23 or the composition containing the same may arrive at the corresponding target tissue. That is, the bacteriophage ΦCJ23 or the composition containing the smae as the active ingredient may be administered through various oral or parenteral routes. Non-restrictive example of the administration route, oral, rectal, local, intravenous, intraperitoneal, intramuscular, intraarterial, subcutaneous, and nasal administration, or inhalation, etc., may be performed.
[0075] Hereinafter, the present invention will be described in detail through the Examples. However, these Examples are only to illustrate the present invention, and a scope of the present invention is not limited to these Examples.
Example 1
Isolation of Bacteriophage being Infected by APEC
Examples 1-1
Screening of Bacteriophage and Isolation of Single Bacteriophage
[0076] After 50 ml of a sample obtained from feces and environmental samples from areas of a duck farm in Boryeng, South Chungchong Province was centrifuged at 4,000 rpm for 10 minutes, the supernatant was filtered with a 0.45 μm filter to prepare a sample solution, and then a soft agar overlay method was performed using the prepared sample solution. The soft agar overlay method is a method of observing a lytic action of bacteriophage using host cells growing in top agar (attached onto a solid medium using 0.7% agar).
[0077] In detail, 18 ml of sample filtrates was mixed with 150 μl of a shake culture solution (OD600=2) of APEC(E10-4) obtained from College of Veterinary Medicine in Kunkuk University and 2 ml of 10×LB medium (tryptone 10 g/l; yeast extract 5 g/l; and NaCl 10 g/l) and cultured at 37° C. for 18 hours. Then, the culture solution was centrifuged at 4,000 rpm for 10 minutes, and the supernatant was filtered using the 0.45 μm filter. Then, after a mixed solution of 3 ml of 0.7%(w/v) agar and 150 μl of the shake culture solution (OD600=2) of APEC(E10-4) was poured and hardened on to a LB plate medium, 10 μl of the sample solution was dropped thereon, followed by culturing at 37° C. for 18 hours. Then, it was confirmed that a plaque was formed.
[0078] Since it is known that one kind of bacteriophage is present in a single plaque, a single bacteriophage was intended to be isolated from the formed plaque. In detail, the plaque was added to 400 μl of a SM solution (NaCl 5.8 g/l; MgSO47H2O 2 g/l; 1M Tris-Cl (pH 7.5) 50 ml) and left at room temperature for 4 hours, thereby obtaining a bacteriophage solution.
[0079] Thereafter, 100 μl of the bacteriophage solution was mixed with 12 ml of 0.7%(w/v) agar and 500 μl of the shake culture solution (OD600=2) of APEC(E10-4), followed by performing the soft agar over lay method using a LB plate medium having a diameter of 150 mm. The culturing was performed until APEC was completely lysed. After the culturing was terminated, 15 ml of the SM solution was added to the LB plate medium and left at room temperature for 4 hours, thereby obtaining a bacteriophage solution.
[0080] After the solution was recovered and 1% (v/v) chloroform was added thereto, the mixture was mixed for 10 minutes, followed by centrifugation at 4,000 rpm for 10 minutes, thereby obtaining a supernatant. The obtained supernatant was filtered with a 0.45 μm filter, thereby obtaining a final sample.
Examples 1-2
Large-Scale Culture and Purification of Bacteriophage
[0081] The bacteriophage obtained in Example 1-1 was cultured at large scale using APEC (E10-4), and then the bacteriophage was purified therefrom.
[0082] In detail, after APEC (E10-4) was shake-cultured, an aliquot of 1.5×1010 cfu was centrifuged at 4000 rpm for 10 minutes and then resuspended in 4 ml of the SM solution. The bacteriophage of 1.5×106 pfu was inoculated thereto (multiplicity of infection (MOI)=0.0001), and left at room temperature for 20 minutes.
[0083] Thereafter, the solution was inoculated into 150 ml of the LB medium and cultured at 37° C. for 6 hours. After the culturing was terminated, chloroform was added at an amount of 1% (v/v) of a final volume and stirred for 20 minutes. Then, restriction enzymes DNase I and RNase A were added so as to have a final concentration of 1 μg/ml, respectively, and the solution was left at 30° C. for 30 minutes. Then, NaCl and polyethylene glycol (PEG) were added so as to have final concentrations of 1M and 10% (w/v), respectively, and further left at 4° C. for 3 hours, followed by centrifugation at 4° C. and 12,000 rpm for 20 minutes, thereby obtaining precipitates.
[0084] The obtained precipitate was suspended in 5 ml of SM solution and left at room temperature for 20 minutes. Then, 4 ml of chloroform was added thereto and stirred, followed by centrifugation at 4° C. and 4,000 rpm for 20 minutes, thereby obtaining a supernatant. Thereafter, the supernatant was filtered with a 0.45 μm filter, and ultracentrifugation (35,000 rpm, 1 hour, 4° C.) using a glycerol density gradient method (density: 40%, 5% glycerol) was performed, thereby purifying the bacteriophage.
[0085] The present inventors designated the bacteriophage obtained by extracting the sample from the feces sample of the farm and having the specific bacteriocidal activity against APEC as "Bacteriophage ΦCJ23" and deposited the bacteriophage at Korean Culture Center of Microorganisms (361-221, Hongjedong, Seodaemun-gu, Seoul, Korea) under deposition number KCCM11365P on Jan. 30, 2013.
Example 2
Examination on ΦCJ23 Infection of APEC
[0086] In order to confirm whether or not the bacteriophage ΦCJ23 purified in Example 1 has a lytic activity on E. coli strains other than APEC(E10-4), cross infection with other E. coli species was performed.
[0087] In detail, among wild-type E. coli strains obtained from College of Veterinary Medicine in Kunkuk University, two kinds of APEC strains (E10-4 and E09-35) and six kinds of non-pathogenic E. coli strains(E09-1, E09-10, E09-13, E09-14, E09-15, and E09-16) were cultured, respectively, thereby obtaining culture solutions. Each of the culture solutions and the purified ΦCJ23 were used to perform the soft agar overlay method, and whether or not a plaque was formed was confirmed.
[0088] The results were shown in the following Table 1.
TABLE-US-00001 TABLE 1 Strain name Plaque formation APEC (E10-4) ◯ APEC (E09-35) ◯ E. coli (E09-1) X E. coli (E09-10) X E. coli (E09-13) X E. coli (E09-14) X E. coli (E09-15) X E. coli (E09-16) X
[0089] As shown in Table 1, it may be appreciated that the bacteriophage ΦCJ23 purified in Example 1 did not have the lytic activity on the non-pathogenic E. coli strains.
Example 3
Observation of Morphology of ΦCJ23
[0090] The bacteriophage ΦCJ23 purified in Example 1 was diluted in a 0.01% gelatin solution, and then fixed in a 2.5% glutaraldehyde solution. The fixed bacteriophage was dropped onto a carbon-coated mica plate (ca. 2.5 mm×2.5 mm), adapted thereto for 10 minutes, and washed with sterile distilled water.
[0091] A carbon film was mounted on a copper grid, stained with 4% uranyl acetate for 30 to 60 seconds, dried, and investigated using a transmission electron microscope (JEM-1011, 80 kV, magnification ×120,000 to ×200,000) (FIG. 1).
[0092] FIG. 1 is an electron microscopy photograph of the bacteriophage ΦCJ23. It was judged that the bacteriophage ΦCJ23 has a morphotype with icosahedral head of a size about 40 nm without a tail, such that it morphologically belongs to Podoviridae.
Example 4
Genomic DNA Size Analysis of ΦCJ23
[0093] Genomic DNA was extracted from the bacteriophage ΦCJ23 purified in Example 1.
[0094] In detail, 20 mM Ethylenediaminetetraacetic acid (EDTA), 50 μg/ml proteinase K, and 0.5% (w/v) sodium dodecyl sulfate (SDS) were added to a culture solution of the purified bacteriophage ΦCJ23, and left at 50° C. for 1 hour. Then, an equal volume of phenol (pH 8.0) was added and stirred, followed by centrifugation at room temperature and 12,000 rpm for 10 minutes, thereby obtaining a supernatant.
[0095] The supernatant was mixed with an equal volume of PC (phenol: chloroform=1:1) and centrifuged at room temperature and 12,000 rpm for 10 minutes, thereby obtaining a supernatant. The supernatant was mixed with an equal volume of chloroform and centrifuged at room temperature and 12,000 rpm for 10 minutes, thereby obtaining a supernatant. The obtained supernatant was sequentially mixed with of 3M sodium acetate to be 10% (v/v) of total volume and a double volume of cold 95% ethanol, and left at -20° C. for 1 hour.
[0096] Subsequently, centrifugation was performed at 0° C. and 12,000 rpm for 10 minutes, and the precipitate was obtained by removing the supernatant. Then, 50 μl of Tris-EDTA (TE) buffer (pH 8.0) was added thereto to dissolve the obtained precipitate. The extracted DNA was diluted 10 times, and a concentration was measured by measuring absorbance at OD260.
[0097] Next, 1 μg of DNA was loaded onto 1% pulse-field gel electrophoresis (PFGE) agarose gel, and for 20 hours using a BIORAD PFGE system program 7 (size range: 25-100 kb; switch time ramp: 0.4-2.0 seconds, linear shape; forward voltage: 180 V; reverse voltage: 120 V) (FIG. 2).
[0098] FIG. 2 is a pulsed field gel electrophoresis (PFGE) photograph of the genomic DNA of the bacteriophage ΦCJ23, and it may be confirmed that the genomic DNA of the bacteriophage ΦCJ23 has a size of about 71 kbp.
Example 5
Protein Pattern Analysis of ΦCJ23
[0099] 15 μl of purified bacteriophage ΦCJ23 solution at a titer of 1010 pfu/ml was mixed with 3 μl of a 5×SDS sample solution, and heated for 5 minutes. Then, the 15% SDS-PAGE was performed (FIG. 3).
[0100] FIG. 3 is an electrophoresis photograph showing a result of SDS-PAGE performed on the bacteriophage ΦCJ23, and main proteins having sizes of about 40 kDa, 43 kDa, and 53 kDa were observed.
Example 6
Genetic Characteristics Analysis of ΦCJ23
[0101] In order to confirm genetic characteristics of the bacteriophage ΦCJ23 purified in Example 1, DNA of the bacteriophage ΦCJ23 was analyzed using a FLX titanium sequencer (Roche), which is a gene analysis apparatus. The genes were assembled at Macrogen Inc. using GS and de novo assembler software (Roche). Sequence analysis of an open reading frame was performed using GeneMArk.hmm, Glimmer v3.02, and FGENESB software. Identification of the open reading frame was performed using BLASTP and InterProScan program.
[0102] The genome sequence of the bacteriophage had various similarities with that of the existing reported bacteriophage, but it was confirmed that there was no bacteriophage of which all of the fractions were completely (100%) equal. Therefore, it may be confirmed that the bacteriophage was a newly isolated bacteriophage.
[0103] The following Table 2 shows results obtained by comparing homologues of the genome sequence of the bacteriophage ΦCJ23 and decoded genome sequences of other bacteriophages.
TABLE-US-00002 TABLE 2 Query Subject Score Identities Pct. Name Length Start End Description E-Value Match/Total (%) contig00001_orf00004 183 1 165 hypothetical protein gp2.4 2E-23 51/55 92 [Escherichia phage vB_EcoP_G7C] contig00001_orf00002 438 1 435 RNA polymerase RNAP1 3E-74 142/146 97 subunit A [Escherichia phage vB_EcoP_G7C] contig00001_orf00001 324 1 321 hypothetical protein gp1 8E-46 94/107 87 [Escherichia phage vB_EcoP_G7C] contig00001_orf00005 129 1 126 hypothetical protein gp6 1E-12 37/42 88 [Escherichia phage vB_EcoP_G7C] contig00001_orf00017 1218 1 1215 RNA polymerase RNAP2 0 358/406 88 subunit A [Escherichia phage vB_EcoP_G7C] contig00001_orf00011 315 1 312 hypothetical protein gp11 5E-45 84/104 80 [Escherichia phage vB_EcoP_G7C] contig00001_orf00008 222 1 219 putative phage protein 4E-32 64/73 87 [Escherichia phage vB_EcoP_G7C] contig00001_orf00006 309 1 303 hypothetical protein gp9 2E-45 93/101 92 [Escherichia phage vB_EcoP_G7C] contig00001_orf00015 819 1 816 RNA polymerase RNAP1 9E-152 261/272 95 subunit B [Escherichia phage vB_EcoP_G7C] contig00001_orf00009 258 1 252 hypothetical protein gp9.2 3E-25 59/84 70 [Escherichia phage vB_EcoP_G7C] contig00001_orf00013 351 1 348 hypothetical protein gp14 1E-60 107/116 92 [Escherichia phage vB_EcoP_G7C] contig00001_orf00019 879 46 876 capsid decorating protein 4E-119 220/277 79 [Escherichia phage vB_EcoP_G7C] contig00001_orf00012 381 1 378 hypothetical protein gp12 1E-61 112/126 88 [Escherichia phage vB_EcoP_G7C] contig00001_orf00018 183 76 180 hypothetical protein gp16.1 5E-08 25/35 71 [Escherichia phage vB_EcoP_G7C] contig00001_orf00020 189 1 186 hypothetical protein gp17.1 4E-28 61/62 98 [Escherichia phage vB_EcoP_G7C] contig00001_orf00023 186 1 183 hypothetical protein gp17.2 1E-24 58/61 95 [Escherichia phage vB_EcoP_G7C] contig00001_orf00027 507 1 504 dCTP deaminase [Escherichia 5E-87 157/168 93 phage vB_EcoP_G7C] contig00001_orf00025 1053 1 1050 hypothetical protein gp24 0 331/350 94 [Escherichia phage vB_EcoP_G7C] contig00001_orf00026 1176 1 1173 hypothetical protein gp25 0 374/391 95 [Escherichia phage vB_EcoP_G7C] contig00001_orf00029 279 91 219 putative membrane immunity 6E-17 41/43 95 protein [Escherichia phage vB_EcoP_G7C] contig00001_orf00031 444 1 441 hypothetical protein gp29 6E-70 133/147 90 [Escherichia phage vB_EcoP_G7C] contig00001_orf00032 945 1 942 thymidylate synthase [Escherichia 3E-148 261/314 83 phage vB_EcoP_G7C] contig00001_orf00034 213 1 210 hypothetical protein gp31 2E-33 67/70 95 [Escherichia phage vB_EcoP_G7C] contig00001_orf00037 2568 1 2562 rIIA-like protein [Escherichia 0 734/854 85 phage vB_EcoP_G7C] contig00001_orf00035 327 1 321 hypothetical protein gp32 2E-25 79/108 73 [Escherichia phage vB_EcoP_G7C] contig00001_orf00045 306 1 297 hypothetical protein gp40 1E-46 93/99 93 [Escherichia phage vB_EcoP_G7C] contig00001_orf00043 531 1 528 hypothetical protein gp38 4E-92 168/176 95 [Escherichia phage vB_EcoP_G7C] contig00001_orf00041 351 1 348 hypothetical protein gp36 8E-56 108/116 93 [Escherichia phage vB_EcoP_G7C] contig00001_orf00048 981 1 978 hypothetical protein gp42 1E-176 316/326 96 [Escherichia phage vB_EcoP_G7C] contig00001_orf00044 2580 1 2577 DNA polymerase [Escherichia 0 830/859 96 phage vB_EcoP_G7C] contig00001_orf00051 804 1 687 ssDNA-binding protein 1E-110 215/229 93 [Escherichia phage vB_EcoP_G7C] contig00001_orf00050 753 1 750 hypothetical protein gp44 8E-138 246/250 98 [Escherichia phage vB_EcoP_G7C] contig00001_orf00042 1311 1 1308 DNA helicase [Escherichia 0 423/436 97 phage vB_EcoP_G7C] contig00001_orf00040 396 1 393 hypothetical protein gp35 1E-61 119/131 90 [Escherichia phage vB_EcoP_G7C] contig00001_orf00039 2049 1 2046 rIIB-like protein [Escherichia 0 576/683 84 phage vB_EcoP_G7C] contig00001_orf00055 123 1 120 hypothetical protein gp47.2 5E-13 34/40 85 [Escherichia phage vB_EcoP_G7C] contig00001_orf00053 441 1 294 hypothetical protein gp47 2E-25 73/103 70 [Escherichia phage vB_EcoP_G7C] contig00001_orf00049 2154 1 2151 DNS protein [Escherichia 0 695/717 96 phage vB_EcoP_G7C] contig00001_orf00047 498 1 495 putative HNH homing 1E-90 160/165 96 endonuclease [Escherichia phage vB_EcoP_G7C] contig00001_orf00052 555 1 552 hypothetical protein gp46 1E-98 181/184 98 [Escherichia phage vB_EcoP_G7C] contig00001_orf00063 444 1 441 putative structural protein 1E-47 109/153 71 [Escherichia phage vB_EcoP_G7C] contig00001_orf00065 843 7 840 putative tail protein [Escherichia 5E-136 285/278 95 phage vB_EcoP_G7C] contig00001_orf00061 10713 2713 10710 virion RNA polymerase 0 2410/2686 89 [Escherichia phage vB_EcoP_G7C] contig00001_orf00071 345 1 342 hypothetical protein gp58 2E-57 113/114 99 [Escherichia phage vB_EcoP_G7C] contig00001_orf00070 1221 1 1218 hypothetical protein gp57 8E-170 386/406 95 [Escherichia phage vB_EcoP_G7C] contig00001_orf00066 642 1 639 hypothetical protein gp55 5E-102 206/213 96 [Escherichia phage vB_EcoP_G7C] contig00001_orf00064 2655 1 2652 hypothetical protein gp53 0 810/887 91 [Escherichia phage vB_EcoP_G7C] contig00001_orf00059 519 37 513 gp49 [Enetrobacteria phage N4] 2E-85 144/159 90 contig00001_orf00069 1203 1 1200 major coat protein [Escherichia 0 389/400 97 phage vB_EcoP_G7C] contig00001_orf00072 2271 1 2268 portal protein [Escherichia 0 735/761 96 phage vB_EcoP_G7C] contig00001_orf00005 348 1 342 hypothetical protein gp71 1E-51 99/114 86 [Escherichia phage vB_EcoP_G7C] contig00001_orf00002 1590 1 1587 terminase subunit A [Escherichia 0 521/529 98 phage vB_EcoP_G7C] contig00001_orf00074 636 1 633 N-acetylmuramidase 4E-103 199/211 94 [Escherichia phage vB_EcoP_G7C] contig00001_orf00073 225 2 169 putative Rz/Rzl spanin protein 5E-24 55/56 98 [Escherichia phage vB_EcoP_G7C] contig00001_orf00075 330 49 321 hypothetical protein gp63 2E-21 66/91 72 [Escherichia phage vB_EcoP_G7C] contig00001_orf00001 711 1 708 putative tail protein [Escherichia 4E-134 232/236 98 phage vB_EcoP_G7C] contig00001_orf00003 690 1 687 hypothetical protein gp69 5E-114 223/229 97 [Escherichia phage vB_EcoP_G7C] contig00001_orf00004 279 1 276 hypothetical protein gp70 3E-27 69/92 75 [Escherichia phage vB_EcoP_G7C] contig00001_orf00006 315 1 312 hypothetical protein gp71.1 2E-43 84/104 80 [Escherichia phage vB_EcoP_G7C] contig00001_orf00004 2115 64 2112 tailspike protein [Escherichia 0 510/689 74 phage vB_EcoM_CBA120]
Example 7
Stability Test of ΦCJ23 Depending on pH
[0104] In order to confirm whether or not the bacteriophage ΦCJ23 may have stability in a low pH environment in stomach, stability test was performed over a wide pH range (pH 2.1, 2.5, 3.0, 3.5, 4.0, 5.5, 6.4, 7.5, 8.3, 9.2 and 11.0)
[0105] For test, various pH solutions (sodium acetate buffer (pH 2.1, pH 4.0, pH 5.5, and pH 6.4), sodium citrate buffer (pH 2.5, pH 3.0, and pH 3.5), sodium phosphate buffer (pH 6.9 and pH 7.4), and Tris-HCl solution (pH 8.2, pH 9.0, pH 9.8, and pH 11.0)) were prepared at a concentration of 0.2 M, respectively.
[0106] After 90 μl of each of the pH solutions was mixed with 10 μl of bacteriophage solution having a titer of 2.0×1010 pfu/ml so that a concentration of each of pH solution became 1M, each of the pH solutions was left at room temperature for 2 hours. In a control group, 20 μl of the bacteriophage solution (2.0×1010 pfu/ml) was mixed with 180 μl of SM solution and then left at room temperature for 2 hours. Then, the reaction solution was diluted step by step, and 10 μl of each of the diluted solutions was dropped and cultured at 37° C. for 18 hours by a soft agar overlay method, and the titer was measured through the presence or absence of lysis.
[0107] FIG. 4 shows a result of the acid resistance test of the bacteriophage ΦCJ23. As shown in FIG. 4, it may be confirmed that the bacteriophage ΦCJ23 did not lose its activity and was stable in a pH range of 3.5 to 11.0 as compared to the control group.
Example 8
Stability Test of ΦCJ23 Depending on Temperature
[0108] A test for confirming stability against heat generated during a formulating process of the bacteriophage in the case of using the bacteriophage as a feed additive formulation among formulations of the bacteriophage was performed.
[0109] In detail, 100 μl of the bacteriophage ΦCJ23 solution having a concentration of 6.5×109 pfu/ml was left at 60° C. for 30 minutes, and then the solutions above were diluted step by step. 10 μl of the diluted solutions at each step was dropped and cultured at 37° C. for 18 hours by a soft agar overlay method, and the titer was measured through the presence or absence of lysis (FIG. 5).
[0110] FIG. 5 shows a result of a heat resistance test of the bacteriophage ΦCJ23. As shown in FIG. 5, the titer of the bacteriophage ΦCJ23 was not decreased by about 1 log value or more at the time of being exposed at 60° C. for 1 hour.
Example 9
Infection Spectrum Test of ΦCJ23 on Wild-Type Strains of APEC
[0111] Whether or not the bacteriophage ΦCJ23 had a lytic activity was tested on 6 wild-type strains of APEC isolated by College of Veterinary Medicine, Kunkuk University other than APEC (E10-4) used in the experiment.
[0112] In detail, 10 μl of bacteriophage ΦCJ23 solution having a titer of 108 pfu/ml and mixed with 150 μl of a shake culture solution (OD600=2) of each of the strains was dropped and cultured at 37° C. for 18 hours by a soft agar overlay method. Then, whether or not a plaque was formed was observed.
[0113] The results were shown in the following Table 3.
TABLE-US-00003 TABLE 3 Strain name Plaque formation APEC (E09-6) ◯ APEC (E09-11) ◯ APEC (E09-35) ◯ APEC (E10-03) ◯ APEC (E10-04) ◯ APEC (E10-05) ◯
[0114] As shown in Table 3, it may be confirmed that the bacteriophage ΦCJ23 had effective infectivity on APEC (including 0-78 serotype), which is a bacterium causing avian colibacillosis in general poultry farms. Meanwhile, it is known that the O-78 serotype APEC is a strain most frequently found in APEC strains isolated in the general poultry farms.
Sequence CWU
1
1
3160125DNAArtificial Sequencea novel Avian Pathogenic Escherichia coli
specific bacteriophage KCCM11365P 1tatcatcaac tattggagct atcatcatga
ctactcaagc tactgtacgt atgaccgcag 60gaactttact tggtactgtt aactcagctg
ctactactgt tgcagatacc ttcggtacag 120caactaaagc agtaggtatg cttaattcat
atgtaagtac tatggcagag aaacaagcca 180ttcgtactaa attagaaatg catacctttg
ttaataaact ggcagaggaa acagctatga 240ctgagactct gcgtaagaaa agtatcgagg
aattctgcaa ggattcagag aacgctcgaa 300tctataacgc agaatattct aaagtaattg
acatcctgac taaagactaa gtcatctagg 360agactcttcg gagtctcctg ttttgaacat
tagatagata gtcaaaagta gataactatc 420tactttactt ctacttctaa aaaccaagac
gctccgcgtc ttatggatga tgtaaatgca 480tccactaatc attgaacttt tatccttttg
tttgggtgtt ctctccgacg agttcacccg 540taaatcacat aaatctggag agtacaaaat
gtctattcaa aaattcactt tcggtcaatc 600taacgctgct gcttctactg ctaaaactga
caaaccaaaa gctcagttct ggctgaacat 660tggttatgta gctaacgaag gctctgatga
tgagaaattc atctctctgc ctactggtat 720tccactggat actcaggagc cactgcctac
taatagcagc aatgctgact tccgtgctat 780gcgttgtgca cagaacgact tgctggaaca
gttaattgag tatgctcaga acctggaacc 840aggtgaagaa ggtatcataa acctgcaagt
tcaacttcgt cgtgtgaaag cagaggctgc 900tgacatccca gcagacgaga ataaatatgc
tcgtaaactg actttctaat caaccaatag 960actcccttcg gggagtcttt tatttttaac
tagacgataa gctatttcta ctttttgagt 1020atctttagac gaagggtact gaaacagtaa
ccagatagat atagatttat attagaagag 1080gaattaacca tgttagattt catcgctttc
tgtgtaatct tctactttct gggttggaag 1140ttacgtcata aatggttgct tattgctaaa
ctgccattcg ttactatcga gtgtctcatt 1200gctagtatta aacacaagaa agcaatgact
cagtactaca agcaacaagc agaagagttt 1260gcaaagagga accagtgata ttcactaaca
tagatgatgc cattgaagaa tgtatctttc 1320gtaggtatca cactggtgta cagaaacgac
actatggtgt tgtacaactc aatggctatc 1380aaatggttgt aagaattgta cgtaagaata
aaccttttaa ctttatgtgg agtactaaat 1440catgcgtaaa tcattaatca tgggaaccag
agaagacgtg caaaagatga aagaacgtct 1500ggttgctaag aaacagacgt ctgaaccagt
acgtaagatt gttaccttca accattcatg 1560tatcaagtaa ctaggagtcc tccattagga
gggcttctat tttttattcc tgaatagtta 1620gatatgtatt taaataggag aaataacctt
gaatgaaatt gtttgtatac ttcctcaatc 1680agctaagata gctgacagga atatccgtat
ccacttaccc tttccattaa gtaatccatt 1740cactcgtaat gaaagaacca cagtagaaga
tgtagaggta gcttatgaag cttacctccg 1800taatcgtctt attagtgggg acaaactaat
tacagcagag atggaaagga ttgcatcctt 1860tgtaacagat agtacaggca agcctgttgg
cttaattggt acagagtctg atgttaacgt 1920tattcgtaaa atattaatgg aggcattaaa
tggctaaata taaggtaata cgctttgcag 1980ataatgaacg tggtattggt gcagaagtag
agaaaaaaac attctttgga accagtacat 2040gggtaaaagt tagtacacat ggacacagtg
agtgcttcga taaatgggtt aataaagaaa 2100ccggtgtatc cggttatgct gatactaccc
aatcagagat taatgacttc tatactgcca 2160atcacattac taaggaataa acaaaatgtt
gttcagcact aattatggta tttgcccaat 2220ttgcaataaa ggaagaggag ttgctaatca
taagaagtgt agtcgtatat tacagaaaca 2280acggaatcag aaagaatggg ataaggtgct
aaataatcag cataaagaag agaatcaaca 2340gatggctgtt aaagcatcta ctcaacgtat
tcgccgtatt aactatatac aggggtatca 2400gaaatgatta ttgcatatcc tacaggcaag
actgtggaat atatgaagca tactattcaa 2460gtgcctcact gggttaagta tatagcatta
ctacctcgtc aatataagag tgctaatacc 2520tcactaatag gattctctaa aaaacctaag
ctaacagaaa acaatatctg ggtatcctct 2580gggagacaag aagagattgg tttttgtgat
ttgagtattg tcaataataa cgtttaccta 2640acgctggaga aagtagcatg aatattgaaa
attatgaaat cactttagta acagcagaca 2700gtaaaactgt gattaacaaa caattaaata
atgaccctga gatgcttaac tgggttgcag 2760agcaatttgc tgatgttaac tcagcacagg
taaccctacg gaatatgtca ggaaaagtag 2820tagcatttac aggcaaggag ccagtatgaa
aactattcta gtaatccatg attctacgtt 2880tactgatgta gataagatga tgcgtaatat
tgattatgta tctcagacaa gccaagcatt 2940caatgaagag tttactctgt actgcaatgc
tgaatctcct ctggttccta tccttaagga 3000atcaggtcta ccattctcta cagagaactt
cccggaagaa ccagactatg taatctcatt 3060tatctatgac ttacatgatg gttctgaaac
tagtgaatta gctatgaacc agtggcgtag 3120taaacgtcct gtgtttgcat ttcaggtact
taaaccatga agattatgtc attaggtgat 3180gacaccaatg ctttacttgg tgtatctgca
cgtccaatta tcattgttaa taaacatcac 3240ggtaaatccg gtgagtatat tgggcgtggc
tcaccattag gcaatccatt cgtcattggt 3300aaacatggaa ccagagagca agtaattgct
aagtataaag tctggttaca agaacagatt 3360gataaaggta atccagttgt actggatgaa
cttaatcgtc tgggtaataa agccattgat 3420gagaaaggat tagccttaca gtgcttctgt
tatccaaaac catgtcatgg tgaggttatt 3480aaagagaagc tagtaaaggc tatgtataac
tactttgtag aaaatctaaa aggataattt 3540atgaaagaaa tcttagtatt cactactaat
gtacttggac agcataacaa tgctgctgcc 3600aaattagctt ataagaaaca cggtgctcgc
tggggcatgg cctatggtca ttatggtaat 3660agctttgcca ttccagtaaa ggatggttat
ggtaaccgta ttaaagaagg tgctatctat 3720ggcttcattg aaggctttat tgcatatgca
tcttctaatc cacaatggga tttcaaagta 3780atgggagata actttctgga cccatacctg
ttcaataatg tcactggcaa tgtgctattg 3840ccagaagcat ggcataaata cttaggtaat
gcttacaact attggagtta ataatgactt 3900atgaagaact atggtctgct caagtcagag
ccagagcact cacacgacac gatatctatt 3960gtgcattaca gaacgaatta aagagtcgta
ctaagctggg tcacatatcc ggcttagtta 4020aaatatctat gactagccta gtctggcctt
atcagaagaa aagtaatgag tttaatggta 4080atggtctgca tgtacgcatc gactacatcg
gtaatgaaaa cagtattcgt attacctttt 4140ggactaagcg ttagttaatg gctgctccgc
agcttttggt tattaatcac atttaggaga 4200ataagcgatg tactcatcca ttgaacatca
gaagcaactc gaaaaactat tcaataaaaa 4260tcagctcttg cctcgcatga ggaaggaatt
cgaggactcc gaagaaattg actttaaggc 4320attcgctgcc tatttggaaa ttgattataa
gctgctcatt gacgcaatgg tgcagattgc 4380cctacataaa cgtgctgaca tccaaactat
gattggttct ctaatgagtc actcagatga 4440tgctcagtac atcgctgact gtctgtataa
gatggcagag aatgactgct tcaactatga 4500ccctaccatc gacaagttcg gtgttatcta
tgagattagt gaagatgtac agatggaact 4560ggaagcattc cagtacccat tacctatcgt
aagtgctcct aagcctgtta agtgtaaccg 4620tgatactggt tactacgaaa gcagaggtag
catcatactt aagaagaatc atcatgagat 4680ggatgtctgt cttgatcaca tcaaccgcat
gaacaatcaa cgtctctgca ttaactggga 4740cgttgctaac tacgtaaaag actcccgtcc
taacatggat aaacccaaag agggtgaaac 4800ccgtcaggac tatgagaagc gtgttaaagc
ctttgagaag tacagccgta cagctaagga 4860agtaatggag ttagtgacca aagaaggtaa
taacttctca ctggctcaca aatatgacaa 4920acgtgggcgt acatatgcct gtggctacca
catcaactat cagggaacca gttacaacaa 4980agctgtactg gagttcgctg ataaggagtt
agtaaatgaa gactaacatt ggctcatatg 5040agctatgggt taatgaagaa tgcgtttact
ccggtacata tgtgaagtgc ttatactttg 5100aaaagcttta caagcttcaa aaccctgaga
gtaaaaccat tatttataaa ctatctgccg 5160acgtagtaac tggctaatcc atccaataag
caggctctcc atagtgggga gccaagaagc 5220aaaaggaaca taaaatgcaa ctgttaaccg
ctaaacaata cctgaaagta gatatcgcca 5280ataactatgg tctggataaa aagacatggg
atgagcgtat tgcctggttc gatgagaacg 5340aagctaacct gcttaatctg gtagatgaag
ctgaagattc tgcactgttc tatgctggtg 5400taaatgcatg gaaagatatg aaagcaggta
agcctattgg ttatgccgta gctctggatg 5460ctacatcatc tggcttgcaa ttactggctt
gtctgacggg tgaccgctct gctgctgaac 5520tgtgtaacgt agttaattac atgggtgaga
atggtaagcc attacgtcgt gatgcttata 5580cagtcatcta tcacaagatg ttggacatcc
ttggtgaagc atctcgtatt aaacgtagtg 5640acactaagca ggcagttatg actgcgttct
acggctcaga agccaagcct aaagaagtat 5700tcggtgaagg tattcgtctc aagacttttg
agaatgtaat ggaaactgtt gctagtggtc 5760cttgggcatt gaacaagttc ttactgcaat
gtggtaatcc agatgccaac cgttacatat 5820ggatacttcc tgacaacttc cacgctgtca
ttaaggtcat ggttccagaa gtacagactg 5880ttaacttctt aggcaaaccg ttcgacatta
ctcgtatggt tcagggtact gaagagaaga 5940ctcgtatgct ctctgctaac atcacccact
ctattgatgg gatggtggta cgtgagatgc 6000tgcgtcgttg taacttcgac cgtgacttag
tggaagctgt gcgagaactc tgcgatgaag 6060gtccatcaga atatggggag attgcaggta
atctggagaa ggtacaagag ctatggagcc 6120attatgagaa gtctggcttc ttatctttat
ccatcctgga ctatcttgac ccatgtacta 6180ttgcttatgt agaccgtcag gtagtagcag
atatgattga cactctgcct aagaaaccat 6240tccctgtaat gactgtacat gactgcttcc
gttgccatcc taactacggt aatgacctgc 6300gtcgtcagta caatcagatc ctgtctgata
ttgctaagag tgacctactg ggcttcatcc 6360tgtctcaggt actggggcaa gagttctctg
ctggtaagct ggatgacagc ctctggcagg 6420acattcttga aacagactat gcgttgagtt
aataaactag cctcattcct tcgggagtga 6480ggcatatttt ttgcttggag gtaacaacta
tgctaatccc ttttgaagta atctctcagt 6540taatactgat agttttatct gtactggttc
tgtctacaat agttcatgcc cacaaatcta 6600agtttgctta ttactactat ggagcatact
cactaggctg ctttgtaatg gcaggatgga 6660tagcttttgc tatctactgt gtaaacaact
aaccctccta acggagggtt tattttttta 6720catgtcatgt atactaactc tgctatttac
ttatgaggaa attgatatgc caactttgaa 6780agtaggtttt aacaaaacca ctaatgttgc
aactgtactg gatgccagtg gttctatccc 6840aggtggttcc gtagaagtag gaacctttgt
acacccagat gccacttacc ctgatagctt 6900agttatcttc catggtgttc gtgacctgct
gtataaacgc tctgctaaag atccttctaa 6960agaaggcttc tggcctaaca acatcgtgga
tatgcagtct atctctattg atatgaaagc 7020tactccacga ctgactattg ctaccaaact
gcctcgtgta gtctctacta tcgaaggtga 7080agacatcaac tggcacgttg atgtagcagg
tgggaaagca ccatttactt ataaatggca 7140attcaaagct gatactgctg gagcagcatt
cgctgatatt gattcaggtg agaacgaatc 7200cgctgctact gcaacactga ctcttaatga
cgtaacagct acttctgctg gtacttacaa 7260agtgattgtt actgatgcta atggaaccac
ggtagaggat gagtcactat tagctgtagg 7320ctattacgaa gcaagttcac tggtagctac
tcctgattcg ctggctctgt ctgttgctgc 7380tgatacaact gctggtaaga ctgtaacaat
tgttgctatg cctgttggtt catcttctgg 7440tgctttatct atcaagactg ctccagattc
tggtcgtgct acggctacca ttgctggtaa 7500cgtactgaca gttaagccag tggctgctgg
tgatactact tctgtagtag ttaccaatgg 7560tacggtagat ataactatcc ctgttactgt
tgcagaataa gggtattctt tgtttggggt 7620tataaaccct gagataaaac cctcaagttt
gacctcccta ttgaagggag gtcttttttt 7680gacttaagca tcctggagcc aggaaatgta
caaagaaatt gcattcttct cattcttatt 7740aggtggttta ataggtgcag gatttgtagc
tatttctaat acctactttg gttctactcc 7800aaattctgta acgcaaaccc tcaagcatga
atgtgaattg aatattcctc gtaatcaaaa 7860ctgtgtaatg cagttcgtac cggagaagaa
atgaatatcc acaatctaca tttggttgag 7920aaactgtatg aagaatacaa caacaaccga
caacaattag ctagtttgaa gaagtcacca 7980catatggtga aagtatcatt caatggtacg
gatttaggac cacaagcccg tacaagaata 8040cttcctggac tactgtcctt ttatcaaagg
agggttgctg ttttagagaa acgctttgaa 8100catttaggtg tagatttatc accacttcca
gatgaaggag aagagggatg aaagtaacaa 8160atcgttctga gaacaatgag gtaactttcg
gtgatgtgga gcctgctaat ggttttattt 8220ataaccagac agtgtgttta aagattcact
tacctgatgg taaacctgct gctgttgctg 8280tagaaactgg taaatcattc tgtctatctg
caagtacatt cgtaacacca ataaacctgg 8340aaggatatta cctttgaaat taacacagtc
ccaagcaatc ttccttcgta tggttcaagg 8400tggctctgcc accagcaacc gtaataacaa
aaccgcacag tctcttaaga agctgggctt 8460agtacagttc aatgctggtc ttgggtggtc
attaacccct atcggtgtac ttaaactcaa 8520tgaattaaaa ggtaactaaa tgaaaacttt
atttaaaggt attgcagtag ctgcactgat 8580agctttggct ccaaatgcac aagctattga
accagaaccc attttggaag gtgctcaggc 8640ttatctggat gacactcgtg atgcgttcgg
acaaggattc ttcatgggta gtatgatctc 8700ttacatcgag agtactaata actgtgttcc
tgaaggtatt aagtattcgg tcattctgcc 8760taagattgcc aaagtagtta tttatgactc
tgcaatcctc aagatgaaga atacatctca 8820aattgttgtg tattcagtac acaaggcgta
tccctgcact aaatcttaat tagcaattcc 8880attagcttta ggagagtaag taaatggcaa
gcatcgactc cttgaccgta tgtaattcac 8940gtcaagcccg taactttatt atccgtgctc
tgaaagcagg taacgtaccg ttcctgactt 9000catcacctgg catgggtaaa tctgcaatca
ttcgttctat tgcagaagaa tttggtatga 9060aactgataga ccatcgtctg tctacttctg
ccccggaaga cctttctggc ttaccattcc 9120gtaatggtga ccgtgcagag tttatcccat
tcgctgactt attcccgatt gagggggatg 9180aagtaccaga aggttataat ggctggctcc
tgttccttga tgagttcaac tcagctaaga 9240aagaagtagt agctgctgca tacaaactaa
ttctcgaccg tatgactggt cagaaaaaac 9300ttcatcctaa tgtgatgatt gtctgtgctg
gtaacaaagc tactgaccgt gccattgtta 9360atcctctggg tactgcaatg cagtctcgtg
tggttcactt tgaaatggaa cttaacttcg 9420acatctttgt tgaagatgta atgattcctc
aacaatggga tgaacgtctg gttgcatttc 9480tacatgctaa cccaggttat ctacatgact
tcgacccagc tcataagaac aaaacgttct 9540gttgccctcg tacctgggac tttgttaata
aagacctcaa gaaccttcca gaaggtgctc 9600tgcctgatga agattccctg tactacagtg
gatttgttac acctggtaag gctgtagagt 9660ttgttcaatt cactcaggta tataatcgta
ttattacgat tgagaaagtg gtcaaagacc 9720cattgggttg tccactaccg gaagataaca
acctgtgttg ggctactgtt aaccatctag 9780ctaacaaaac tactgaagag aacttcgctg
atgttcttca gtacatcgaa cgctttaaaa 9840cgttcaccca taagattctg tacttccgta
cagtaggcag aacattacca gaacttcagg 9900ctactcctga atggcgtaag gctgctgcta
atatctctcg ttacattcac ggataaaaca 9960atgaaccaat ttcctcagca cacacttagt
gatgaacaac tcatgcgcga atatgaccgt 10020attcaggcgc aggcgtttct cggacgcagt
gctgccttct ttggttcatt actatgtagt 10080cttaaattct catggaaacg tgaggattgt
cccactgcat gtactgatgg gatagaactc 10140catttcaacc cagacttctt tatctggatg
tgtccagatg caagggaaac agtattaatg 10200catgaactat ggcatgtggc atatctacat
gacatccgtc gtggaagccg tgacccggaa 10260gtctggaacc aggcatgtga ccacttcatt
aaccttcagt tagaggagga tggttacaag 10320ttcactggta ttaatgaagg catttgcaaa
gaccctcaat ataaaggatg ggtcgaagaa 10380gacatctacg atgacctgat gaagaaccct
cagaaaaggc agaagccgtc agggggtgct 10440ggagcaggtc ttgctggcga catgaaatcc
cccacttcgg gacagtccca gggtgctgtc 10500gtcaacaacg tagtacgtgc aatgcagagc
cagaagatgg ctggtggaac aatgcccggt 10560aagggtgctg gtcgtatgga agaggttatt
acccaattcc ttaaaccagt ggttccatgg 10620caagaagtac tcatgaactt ctttaccgac
attgatgaca ctcactatac gtgggccaga 10680cctaaccgtc gttacactga catttatcta
ccttccctgg aagatgatga aggacgtcta 10740cgacacctag cctactttga ggatgtatct
ggttctatta gtagtgctga ctctctgcgt 10800tttaactcag aggttgccta cgttaagagt
caattcaatc ctaagaagat gaccctaatc 10860accttcgacg atgttatcca ggaagaaata
gacatcactg aagaagatac tttcgaagag 10920attaagatta ctggacgtgg tggtacaaac
ctggaaccag tacgcgaatg gattattaag 10980aataagccaa ccgctgcaat catattctct
gatatgtatg ttcgtccaat ggaagaattg 11040ccatttgata ttcctatcat ctggtgtgtt
ctgaataatc ctaatgctac cgtacctttt 11100ggggaggtag ttcatatccc taaaggaatg
aaataatggt tgttaatggt aattctctat 11160atcgttcatc tcagttgctg gatgtcccag
accgtaagat atccgagcat ggtgtaagct 11220atggattagg tgaagctggt tatgatattc
gtatcaaaca ggatattacc ttctatcgct 11280tatttgggtt gattccaatg gtgaaggtcg
ttgatagaaa taaagtatca cgccatttcg 11340gcaagttcac attggcttca gcaattgaga
agttcaacat gtccccttcc tgtgtagcta 11400tcgttcacga taaatctaca tgggcaagac
gtgcattatc tgtgttcaat accgtaatag 11460agccaggatg gaaagggtat ctcaccctag
aactggtcta tcatggtcgt aagaaattgc 11520atatcccggc tggtgctggg atagctcaag
tattatttca tctggttcag gaacctgcta 11580attacaatgg caagtatatg aaccaggaaa
accaaccagt agctgctaga tcctcaaaat 11640aaaggactat ccagcttaat caaggaaaca
acatgtcagt atttcaagta actcaagaaa 11700gtacaggtat tcgcctaacc attaatgcga
accaagtgat tgcagtacaa gaacttaccg 11760caggtaactc tgctattact accgtaggag
gtgatgtagt aattaccaaa gaaacctatc 11820gttcagtacg taattacttg aaaaaagctc
ttgctcctgc aagcaaagat gctgagtaag 11880tagctgccta aatagcccag catagttggg
ctatttgtga agaaactaac tcaaccccac 11940ataggaacca tcatggaatc tttagcagca
atccttgttc tgttatttgt attagctgta 12000taccttatcc ctactattat tgcttttgca
cgaggacacg cctctaagtg gggtattggt 12060gtcctaaata ttgtattagg ctggtcttta
gtcttctggg tagtagcact gatttgggca 12120ctgtctaata aaggtcagaa tcaagttaca
aacgtaactg ttgttcaaac caatagtggc 12180agtaaaacag agtaactaac ctaagcatca
ttgcatagtg gtgcttgtgg aagttacttc 12240cagctcattg cttatccatc tcctaaccca
agcccaccta acccgtgggc ttttttattt 12300acaggtactc ttatgtcaag cagacaaaat
ggtaagtcta tcctgcaagg actggactta 12360agtaagctgg aacaaactgc aatgcttact
ttaggtaaaa ccatacatga ccaagtggaa 12420atggatggga ttagttctga tgtctatgca
aaaatgcagg ctcttattca gaaaggatgg 12480cccagcagag tatttaatat acctacatac
attcccccag aaccccatct gaaatctaag 12540gtagaccgaa tcatagataa gttctggctt
aacccatgtg gagatgacat gcatctttac 12600ctggctcaaa tacaaaagaa tcctcgtact
aaggatgtgt ttaaaagtaa gagcactacc 12660catcactacc catggtatag aaggggtagt
aaatactaat gcgtatccca ttcctaagaa 12720aaagggaaca aaatcctgtt ctttataata
aaggcataga ggatgagtat gaactcaacc 12780gtaaagctcg ttcctacaca acaaaattat
ttatggggac taaaacccct gatagattac 12840tggactttgt ttttgaacaa gtctttatca
tatacagctt agctatgtct gctggttcac 12900aagaagtaag tgataaggca agacatgctc
tctgtatgct ccgtaaagag tatgaagccc 12960tcatgtatga agacttgtac tcctttaaag
aagaaaccgc tgtagcatgt tctgtggctc 13020ttactgaagg tgtagcagta ttacaagaac
taccccgtag tgagtttaag ctggtatacg 13080ttcaggttaa gagaatcaca gagacaagaa
gtggtatcac taactatcta aggtcattct 13140aatgattaag gcatcagtaa ttgcagattc
cgttcatcca gaaacaggaa cccgtatcac 13200aacctttgaa ttggtttatc cccgattcat
tcacagtgag tttatgactc accgtgtatt 13260caaccgcaat gcttcaagta gccgtgctat
tcctacctct aagttaatcg aacaggttcg 13320caatgaacca gtgatgccaa gtcactgggg
caagaaccag aaaggtatgc aagcagatga 13380agaactcact cctatggaga ttgaggatgc
taagtttatc tgggataacg ctgcatctgc 13440tgctgctgtg tatgctgaac agctacgccg
tgggcaagta cataaacaga ttgttaaccg 13500tattctggaa cccttcacac atatccgtgt
agtggtaacc tcaactagct gggctaattt 13560ctatggactg cgtgaccaca aagatgcaca
accggagatt cgtgaactgg ctcaagcaat 13620gcgtaaggca cacgaagaaa gcacaccaag
agcattaccg tatgggcaat ggcatttgcc 13680atatattgct catattgacc gtgttggggc
ttacaatttc tgcaaacgta atcgaattac 13740acgcgatgaa ccaagtgatg aagaagtgca
tggactactt ctcaaggtaa gtgctgcacg 13800ctgtgctcgt gcttcctata acaactttga
gggacgtccc tctactattg aggaagacct 13860tggcttattt gctaagttag tggaaaacca
acctattcat gcttccccaa cggaacatca 13920agctacgcct atgaaccttg gtgagaagta
tgtgaataac atgaacccag ttacctggga 13980acaaggtgtt acatccatgg ataaagaagg
gaatctgtac tcaggtaacc tgctccactt 14040tatccaattc cgtaaattaa ttcctggtga
gactattact gaatgaaaaa actagctcta 14100tacgcaatgc taattagtac cctactaaca
ctgacctacg catacaaagt tgcctttgtc 14160gtagaaacag atatgcagtt cattcgagct
agtattctgt tctttgtaag cgagattggt 14220ttatggtgtg tttactactt tgctcgtgac
tacgaagcaa ttcgtgagca agaagaagtg 14280aaaaaacaaa tgatacgatt tgtagaacaa
aatcgtaagt aaacctaaga cctcctccgg 14340gaggtctttt tttggtttca attaactctt
tcaggagggc atatgcctgc taaataccgt 14400atcaaagaca cacccgtaat gtgtgagggt
gagaagggcg acattgtata tgcctgtatc 14460caggatgatt tcaatgctgc tcagatgcta
acccaaatga caaatacact gcatgtgtca 14520gtaacactgg accctaccgg tgactatcca
tgcttcccta ttcctgccca taacctggag 14580caaatccatg attaaccctg aagtaattca
tagtaaaacc ggtaaggctg tcccactcag 14640tgagattgca gtaactggtg atattgctgc
ttgtccagct aacattgcct ctctatgcat 14700atgcatcgct gcactagcag aggaacgtaa
gttatggctg gaaccaagca aggaaatgat 14760tcaggctggt ttagctgaag tacaaaacac
gttagataac tgggaagaga acggcccact 14820accatatgga acggtcaacg atataacaga
tgacatggca tcagatatgg ctgtgtttgt 14880attacaagca atggcaggta aacgcaatgg
ctaatgtaaa tatcgcctca gagaaaacgt 14940atagcattca gattaatggt ttaaccgagt
atcaggtctt attcctaatg aatgcttttc 15000aaaatagtcc tgtaggtcac catcctaatg
atgaaccacg ggaagaagct gaacttcgta 15060aagctatttt tgataagtgt aaacaagttc
taatgtaagc aacatttaat ctggagagta 15120aataaatgtt agttgcagat accaacgaaa
tagctacctc agcgacactg ggtggcaaag 15180aaacaattgc ctttggcatc tcagatgact
cggcattctt ccatgtatta agtacttccc 15240tgtataacaa tcctactctg gcagtagttc
gtgagactat atgtaacagc tgggatgctc 15300atattgaggc aggtaaaact gataccccta
ttcgtatcac cattgataca gacaacttta 15360ttaccttccg tgattacggt agtggtattc
cagatgaact cattggttcc atttatggtg 15420tctatggtgc atctactaag aaagccaaca
ttagtgttac tggtggcttt ggtctgggat 15480gtaaatctcc attcgcttat acagatagct
tacaggttac ttcatggaac caaggaaaga 15540tgtctgtata taacgtagct aaggctgcga
ttgagaatga tggtaagccg ggtattgtcc 15600ctattgttac caatatacct actgaggaat
ccggtctgga agttaaattc cagttaggca 15660aacatgattt aaataccttt attcattaca
tcaagtcaat cgtatttaac ggtgagatta 15720aagctgagct tagtatccct aaactcgtta
aaacggaaga aggtaatagt attcaacaag 15780gtgactacac tttactgaat acgctgggca
tgtcatttga acctggttca tatgatatgt 15840ctgatagatg gtatcagggc tatatgggta
gcagtaacat atacgttcgc tacggtaatg 15900taatgtaccc aattgtatcc agcccagcta
gtgaagaagc tgtaggtctt atcctcaact 15960tcatgaatat tattggtgct gacaatttag
tagttcaggc tgcaccagac accttagcta 16020ttgctcctag tcgagaaaca ctgtctaacc
agaagttaac cgacgatggt attactactc 16080tatgcgtaga tttagtagac cgtatggaga
aagagattaa ggctaagatt cctgaagcca 16140ttaagcaggt tgaagaatat gcctctgaat
cctctactcg cttttgggaa tatccatctt 16200tcttgggtgc tgttacagat agaactgttc
aacgctatat gtcttctagt ttatggacta 16260aacaacgtaa gcatcacata aagcactggc
gtaacttatc caataaggcg ttattagctc 16320gtcctgaata tgcaggtctt aagaagctgt
atggtaaggc tatgcatgct cttaaggata 16380ctcgtgagga aagtacatac tccccattct
cagaattagt atatcgccat ctgcatttac 16440ctcgacttgc tgctttaaaa acctctggta
ttaagtggtc tggttacata atgaatcaag 16500gcaaccgtgt tgacttggta aaaggtaaac
ttactgacta ctttaggata tacaataact 16560ctcaccaaag tattggcata tttactacca
agaacgttgt agttactcga cgtttgtctg 16620attgtgcaga ctctttctca tacttccctg
aatacaatcg aggtgacctg gagcgtacag 16680cttttgttca tgtcgttggt cctaagaaag
gtgcagcaga ggaagctgta gctaagttca 16740ctgctatggg ttatcgggtg attgacctta
ctcaatataa tgagtgggac aaaccaacta 16800acttccgtag ggaacaggct aagatagctg
ctgagaaacg ggctaagaca atagctgtca 16860ataaaaccaa agcaggaggt aaaactaatg
ctctgatttc attgaatgca gttcttggtg 16920ctacccaggt acgtaataaa aatggggatt
gggaacccag gccgtatatt cagaaagaat 16980ttgctgaccc aagtcgtcat gagaaatatg
gtttcgttga gatagaacag cctaagtact 17040acgtactagc caatcaggtt ggttctggta
gtcctgtaac tgctcgaatt ggaaccatgt 17100ggaaatggta tgagttgcct gatgagatga
aagcagagac tgttgtctgt cgtaatcaaa 17160tcgaagctaa caaggccaaa cgacgaggtg
ctatccacat tgatgatgtt cgctttagtg 17220aattgatgtc tgttattacc agcaaaggct
tcaagaaata tgttactgaa catcgcatcg 17280gtattcttga atacgtagga ctggatgaca
gggaatactg ggaaattcta gacatactgg 17340gtctaacctt caaaccctta cagaatctga
ttttcaaacc agagtatgaa tgggcatatg 17400actttttacg aaatcgacca cacgacaaca
aagaaaaact tgttgaaatg ggttgtatta 17460agtcagttga tgacttagag ccatatgtaa
aactggttaa tccccgtaac cataagtact 17520ttaaagtact taatgattac aaagaattat
tcagttatag ctggaataaa aatgacatcc 17580tacagacgct ggacttaggt agtttggtca
aacaccttaa gaagaatcca gaggatattc 17640ctgggtttaa gtccctctac cgtaatcgtc
taaataaact gaaaggtaac taatctgatg 17700aaaatcgaac tgatttctat catcgcactg
gcagttgaca gtcgtaacct tacattgtgg 17760aagcccgatg gctccacgat tgtatatcct
cagagagacc cacgggttgc tcgcattgta 17820tctgaagctc aaactaaagg tctgggaact
accaaagacc aaatagaagt aaacatcgca 17880ccagaagtaa acctgcgtac tgaatatctg
gaagcagaga agaacactaa cggattcgtc 17940cgtttcttca aggtagctaa agctaaactc
aaagagttct tccaggatgg tacaggtgtt 18000caacctgacc gtattatttc tgatattaag
ctgggtaatc ctactaagac actggtgtct 18060aaagctatgg atacctttct ggctgtacag
gccaatgaac cagaagtaac tgtaacagat 18120ggttgttatg acaaacgtga caatctgatg
tgggttactg gctgggacaa agaccataac 18180catcctgcac tggttcgttt tattaacgac
gtaatgggtt gtgggtatga atatactaac 18240accatgctca aacaggaatg gcctgttgca
gttcgtgcag tatctgatga tgaaatgggt 18300gagtttgcta aacaagccga acacatcaaa
ggggttcatg ttgtattcac cagtcgtaaa 18360cataccccac caccttacat tgaagtaact
aaagttacga accaggataa gctggctgct 18420gcctcggaga aactagctgc attgggtgct
attagtactg atgacgctaa cttccacacc 18480gatgtgaagg aagatgaagt ggttgttgct
gttaccaaca atggggttat ccccggagtt 18540gagaacctgc aacgtcactt acgtcagtct
gccaagctga aagactacaa gggctttact 18600aagttccttg aacgtctggc tccggttatt
aaagaccgtc tgcactcagt agaagacctg 18660atgaagttca tggaaactgc tgaactgcct
attgccgatg atggttcaat cctgttcctc 18720aaacgcctta agtctagtgg tatagaaaac
ggtaaacgtg tattcgttga ctgtcactcg 18780ggtaacattc gtcaatgggt aggctgtaaa
gtgcaggtac gcgaagacct ggtagaccct 18840gaccgtcgtc aggattgctc taacggtctg
cacgtagcat ccatgagcta tctacgtggc 18900ttcggtggta atgtgaccat ccttggtaaa
gtagcaccgg aagatgtatt tgctgttcct 18960cagtacagca ccaataaaat gcgtgtatct
gcatatcata ttattgctga actaccggaa 19020gaggaacgta ataatgttaa taatggtatc
tacctgtcta agacagaagt aggtaagaaa 19080ttgcttaatg atgccatcgt tgggaaccat
agctcaccta ccacacttat tatggttggg 19140ggtcattatg gtactaacct caaatacact
aatctcacat ctggttctgt agaacaattc 19200cgtacagttg ctagcaaaga agcactgaac
atggaagagt cactgaatga agctgtagct 19260gctgaaccag tgaaggctac tgaccttaaa
cctgttatta agaaggctcc tactgtgaaa 19320gaacaaatcc aggaactggt caaagagttc
cttactgcaa caaccccaga agataagtta 19380gctgctgctg accttctggt agaactgcgt
ggtaaagctc gtaagccttg ggctgcattc 19440gatgtgggta atgatgtagt agctaagatt
gctgatgtac gtgctaccta tacagctaag 19500cctattggta aacctaaagc tgttaagcag
gataagacag ttaaacctac taaatctaag 19560cctgctatta atagtactaa tgccaacatc
atcagaggtt atctagcaga tagtggtatg 19620tctgattatc agaaagccca ctccattcat
gacctgaaac gtgcagctaa gaagtcttat 19680gctgctatgg gtcttactga agaagagtgc
aaagccattg ataagctgaa gcaccacctt 19740aagtaatagc ctgttcaaat agcctcactt
ataataaagt gaggctatct ttgaagagga 19800aataagctat gtctaaagta ttcagaagta
atcgcaaagc aactgatgaa gacatcattc 19860gtatgaatgc tgttggtctg tccctcgcaa
ctatcgctaa gacgttgggg gttcacccaa 19920ccacagtcac tttgcgattg cgttctctaa
acattgaacc agccgacaca cgtcggacgt 19980tcatggaaaa cgtattacga cctttaccaa
cccatgtggc tgattggctg tcagaacaag 20040ttggtcctgc ttatgagatt cgctcatatg
taagagactt gattctggag gcatataata 20100atcgccacct taaccaagag agtgagcatg
acaagttcat ccgtttgtac gctggcaaat 20160acggaagcct ggttccggaa agcagtacca
aatccgacaa gtaagaacat tagtacccag 20220atgggttgcc atttggaaga agtagaagaa
atgcttcaga ccatttatcc aaatggtagt 20280tacgatgcag aattactgca acgtgcacag
gatgccatta caaatctggc aaatcatatg 20340aagcgtaaag acaatgccta tcgcattgat
gttagtaccg acctgctgga ctcactggca 20400gaccagattg ttacagcaac tggcgtcggt
actttccttg ggatgaatgt ccctggagca 20460ttggctgaag tcaatcgctc aaactattct
aagtttgaag atggagaacc tgtcttcaat 20520gagaacatga aagttatgaa agggaaagac
tacactcccc cggatttaac cccttacatc 20580taaccctcta cggagggttt tttactggag
attttaatgt tttccaaacc taccaaagcc 20640ccactgaaca aggggcaaga agcggttgcg
aaggagttct tcgacttcct gctcgaccct 20700aatgctaccg aattcaatat tagtggccca
gggggaactg gcaagacatt cctgatgtcg 20760cacctcattg atgacactat gcctgcatat
atggaaactt gctcccttat gggaaccaag 20820cccctatata acgaagttgt tatgactgcg
accacgaaca aggctgctga agttctggct 20880caagctactg ggcgtccaac atctacctat
cattccttcc agggattgat tgttaagaat 20940gactttaaga ctggtgaggc taatgtcgta
ccgtccaaat cattcaatat taagaagaac 21000aaaatcatct tcgtagacga agcatccatg
attgaccgtc agttacttaa atatgctcgt 21060gaaggtactc accagtgcaa actggtattt
gtaggcgatg cttctcagct tctgcctgtt 21120aaagagaata agtctccagt gtatgcaggt
aatatcccaa cacactatct gactgaacag 21180atgcgtaccg atgcaccgga acttaaagca
ttgcaccagc aattgcgtga tacggtagaa 21240ggtaagacag gcttcctgcc tattaaatgt
attccaggca tcattgattg ggtacagggg 21300gaagagatgg agaaactggt tctcagtcac
ttcactcaac ctactaatag ccgtattgtt 21360gcttacacaa atgaccaggt tattaattac
aataactaca ttcgtgaagc taatggctac 21420gtgggtgagt actccattgg tgaacagcta
gtctctaact ctgctattcg cttaggtgtg 21480gataatcgtc tgtctatcga gcaagacgta
aaactcattg accaggatag cagtactcgc 21540atgattccag ttacagatga cctggaactg
gaagttcgtg atagtactct ggaccttggt 21600tatggtggta ttgtaagtga agtcccagta
cctaccgacc cagaatactt caaccgtttg 21660gttaagtggc taggtaaaga gaagaactgg
gaaccctact tccgtcttaa agaaaccatt 21720ccagacctac gtgctactca tgcatgtact
gtccataaat cacaaggctc tacttacgac 21780acaatcttca ttgatgcaga tgacctctca
agctgtcgcc aacctgatat ggttgcccgt 21840ctgctttacg tcgctgtgtc tcgtgcccgt
aagcgagtag tgttttatgg caatcttgtc 21900agtaagtatg gtggtctaac tttctaaggg
aggatatatg cctcagattg gttcagcgac 21960tattggtcag gttgccaata gcagtgagat
agtcaaacac ctgttcttag cagaactggt 22020tcgtcttgat agtgtgttaa acggtatcat
tgataagaac gaccgtatca atggtattga 22080tatatcggct ggatttcttt atcaagggga
gttctatcag cgttctaatg ctgccagacc 22140tccaacctac ggtgaacgat taacacttaa
tccagaactt tggcctgcaa tggacaagta 22200tctgaaagcc tccagtcgcc tgattatgga
agtacacctt gtgaaccaga ctgtatatcg 22260cctggttcgt ggttgtatgt cctatcagga
tgtacgtgat gctttacctg aatgcctggt 22320agcccaggac cagactggta agtacaagga
actgccacgt actcgtgaag cagcctggac 22380acttgctggt gatcctatgg caataaaaca
gtatgagaag attcttccct ctattgagta 22440ctatgcagct tcccatctga ttttctaagg
taaggctatg cgttacatca cctctcagga 22500tacgggtaag tatcctattg ctatcctcgg
tcatcaaatc cgaagggagg agatgattaa 22560aacctacctg ctgcctaatg acctaagcat
ggaagatttc atcttcatcg aacttcattc 22620tgcccccggc aagaagaaga ctcctgcaag
ggagattaag gagttcatac agcaggagtt 22680gcaacaagta ctggacgatg cagagactca
atacattatc tgtaccgatt ctgactactt 22740caaaatactg actaaagaag caaaagcaga
ggctaacctc ggctacgttt gtgattcagt 22800atggggtaag cagaaggtta tctatgcacc
tagctacaga caggtctttt atgaccctcc 22860tgtagtgaaa tctaagattg ctcagggtat
ggatgcatta cttaaccaca tacgtgggca 22920gtatgcagaa ccaggtcagg gaatcattga
gtttgaggct tatccagata ccccagagaa 22980gattaaagcc tggctagacc agttgcttga
gatgaataag ccattggcta tagacatcga 23040ggcattcggt ttaaagcact ataacgcagg
tataggaaca attacgttct gttggagtaa 23100gacacaaggc atagccttta atgtggacta
cgagccgatt cctggagcta ctgaagcacc 23160atacgggcgt atcaacagaa atgatgttgt
tcgaaatctt cttcgtgagt tcttcattaa 23220gtacactcaa cggcagatgt atcacaacat
tagctacgac gtgtatgtgc ttatctatca 23280gttattcatg gataacctga ttgatacaga
aggcttactg catggtatgg aaatcatgct 23340acgcaactgg gactgtacta agttaatcac
ctacctggct actaacagtt gtgctggtaa 23400tcaccttagt ctaaaagacc aggctcagga
gtatgctggt aactatgctc aggatgacat 23460taaagacatt cgtcttattc ctaatgagca
actcttacgt tacaacctca ttgatggttt 23520atgtacctgg tacacctatg agaaacactg
ggatactctc attgctgatg accaactaga 23580tgtttacaac aacatcttta agccagcctg
tgaagatatt atccagatgc agttaactgg 23640tatgcccatg aatatggata ccgttaacca
agtagctaag gagatggaaa ctgacaggaa 23700ccaggctctg aaaactattc gtgagtctaa
gctcatgaag aactttaccc tgatgcttcg 23760tcaggaatgg gtagatgata agaatgctaa
gctcaagaag aagcaggtaa cacttgctga 23820ctgtgatatc gagtttaatc ctaactccgg
tccacaacta cagaagctat tatttgacta 23880tattggctta ccggttcttg gtcttactaa
gagcaagcaa cctgctactg acggtgacac 23940tattaaagca ctgcgtacac acacgcagag
cgaagatgtt aaggaactgc tcaatgcact 24000tatcgactat aagctcgtgg ataagattat
cacttcattc atcccggctt tccgtaatgc 24060acaaccggga ccagatggat ggcactacct
attcggcaac ctcaatctgg ggggaacggt 24120ttctggtaga ttatctgcct ctgagccaaa
cctgcaaacc attccgtctg gctccaaata 24180cgccaagaag attaagaaat gcttcgaagc
acccccaggt tggatctttt gtggactgga 24240ctttgcaagc cttgaggacc gtatctcagc
tttaactact aaagacccta ataaattgcg 24300tgtgtatact gacgggttcg atgggcactc
cctcagagct aaattttatt tcggcgagca 24360aatgccggat atagatgatt ctgtggaaag
catcaactct attcagaaaa aatataaagc 24420cttacgtagt gaatcgaaag ctcccacttt
cttattgact tacggtggga cttatatggg 24480cttgatgaaa aactgcggtt tcccggaagg
gaaggctaag ttaatcgaat ccagatacca 24540tgaaatgtat acggttagtg atgcctgggt
tcaagctaag ctagacgatg ctgccaaaac 24600tggttatgtt actgccgcat tcggtttgag
agtgcgtact cctttactgg ctcaagtatt 24660acgtgggaca tgtaagactc cgtatgaagc
agaagcagaa ggcagaactg ctggtaatgc 24720tttagggcaa agctggtgtc tactaaataa
ccgtgctggt tcagagttta tgcgtaaagt 24780cagagccagt gagttcaggt tagatattcg
tcctagtatt catattcatg atgctcagta 24840cttcatgatt cgtgacaaca tggatacttt
gcaattcacg aacaagcact tggttgaagc 24900cgttaactgg caagaccatc ctgatattgc
tcacccagaa gttggtttgg gtggggaact 24960atccttgttt tacccaacgt gggctaacga
gattgaaatt ccaaatcacg ctaccccaga 25020agaagttcat caaataattc aaaaggcatt
cgcatgacca aaagtactaa agaaactgtt 25080gtcagaaaat atcattggat ggtagcagca
caggtagtat tccaacttcc taaagtggat 25140gatggttccc tgcttaccat gaacacaatg
ttgctcacag atgaacctta cgtgacctat 25200aaagatttgg ctcgtgccaa tcactctctg
aaaatcagtc tggaccagcg tttcgacact 25260tcagttgact tgaaagacat cgtttatctg
tctcttagca acctgggtct gatgtctgaa 25320ccagagttcc aggcaaacat gattcccaag
gagaaataat ggctaagctc tccggtggat 25380tgaataactg gtatgtagta ccagttaagc
accctcaacg gaaagagcaa gagccatacc 25440aagcagagtg tgaggatatt atccaagcac
tgggcatgac cttcgatgaa ggttgtgcct 25500ttaaagctct atggcgaaat gctgctgccc
gtatgggtaa tggtaaacct ggaaacactg 25560ctgtttacga tgcagagaag ctggttcatt
atgctaatcg tattcttgct aaggagaagt 25620tagctagtga gttatttccg gatcctgcta
cgaatgtaaa taccgttggt agttattggc 25680accatacaaa caatggtaaa cctcacttca
ctaaagaatt ttcattcatt gagattgttt 25740ataaagatga acgagatgaa atttactcgt
ataacttcaa tcagctaagt gaaattaagt 25800ggaactgggt tcacagatac aggattactt
actaatgaag ataaccaaca accatgatgt 25860atcactggcc ctggctgtat ggctattgca
tgacgagtaa ctaattatga aacagaggat 25920tgcccatgac aggcttttac agcttgttag
ttatgaccct atttccggga tttttactcg 25980taggaatacc ggaaaggtat ctggttacct
aatgaagagt ggttacgttc aactccgtgt 26040ggatagtgtg ttgtactatg ggcatatcct
tgcatggttc tatgtgcacg gtgtatggcc 26100tacggataga attgaccata aggacaatat
tcgccatcac aactggatag ataacctcag 26160agaagcgacc cacaagcaga ataaccagag
tgctgtttta tctaaaacaa acacatctgg 26220atttaagggg gtatcctttt caaagaattt
aggtaaatac agagcaacta tttgggttaa 26280cagtaaacca attacattag gttttacaga
tgacccaaga gaagctgctg ttctctatga 26340tgaagctgct ataactcatt atggtgagtt
tgctaaaact aataagcaat tgggactgtt 26400atgaaactta ccaacaaaca cgacgttagt
cttgcactag ctgtatggct tgtaacggat 26460gattatgatt atgtagacaa tcctaagtat
ctgtctgtta ctaccttgct taagcccatt 26520aagcaaatag tcatgaagca tcgtgtagat
cttagtgacc agtcaattga tgttatggat 26580ttcgtctcca catcaatggg tactggttta
catgattcta tcgagaaggc ctggaagctg 26640ggtcataaga ctgcattgaa aaagttgggt
tatcctcaac gagtaattga tgcagtagtc 26700attaacccaa ccaaagcaga ctttgatgct
aaccctgacc ttatcccaat ctacattgaa 26760cagcgtggaa ccaggatagt taagggttgg
actatcggtg gtaagttcga catcgtaaca 26820gaaggtctgt tgcaggactt taagtctacc
tcaacctatt cctgggttgc tggttcccgt 26880gatgatgaac ataagatgca aggcagcttg
tatcgttgga ttcacaacga catcattacc 26940gaagatgtaa tccgtattaa ctacatcttc
actgacttca tgaaacacat ggctaatagc 27000aatccgaact atcctgctaa tcgtattatg
cataaggata ttccgttgct atctgtcgag 27060aaaactgaac gttgggtaga agagaagatt
cacctcattg aaaagtactg gaatgcacct 27120gaagaggaaa ttcctgaatg tactgatgag
gagttgtggc gaacagagcc acagttcaaa 27180tacttctctg atgcttctaa ggtagatgta
cctggagcca gaagtaccaa aaaatttgac 27240gatatggcat ctgctcgtat cttcatggct
gaaaaaggtg gcaagggtgc tatcaaggtc 27300gtggaggggc aggttaagcg ttgtctatac
tgccctgtcg cgtccatttg caaacaaaga 27360gagagatatt ttccatcatg agtattgacc
tgaccggagt cactcaccac cctgcaattg 27420aagaaattgt agacgtgctg tgtaacaaga
cacaaaacaa cgacagagga ttcttccgtg 27480tcgaagtagc ctacttcctg gctaaaatgg
catcctgcat gggtgcaacc attgtcacta 27540aagaccgtgg tgacttacca gtcaacattt
acgctatggc attagcaacg tctggcttcg 27600gtaaaggtca ctcggtaaat attattgaag
acggcttcat gactggcttc cgtaaacgtt 27660ttatggaaga caccatgccc gtcattgcaa
atgaccgttt atggaagatt gctaacgaac 27720gtgctgctcg acaaggtaca gaccagaatg
atgagtttga taaagtcgaa gcagagtata 27780aacgtgctgg agcatatccg tttacgtttg
actctggtac tccaccagca gttaaacagc 27840tacgacataa gctgttaatg gctgggtgtg
gttcaatcaa cctacagatt gatgaaattg 27900gttcaaacct gttggctaac acggatgtat
taactctgtt cctggaatta tatgaccagg 27960gtaaggttaa acagaagtta accaagaaca
ctgctgaaag tgttcgtggt gaagaactgg 28020atggtaagac tccagctaac ctgttgctgt
ttggcacgcc aagtaagcta ctagatggtg 28080gtcagaccga agaccagttc tatgactttc
tggatacagg gtatgcacgt cgttgcttat 28140ttgccattgg gcatttagat aaacgagcac
atgcaacaat gtccccagaa gaaatctacc 28200gtaacctgat taagcaggat aacgtacagt
ctctgggtaa gtgggctaat cacttccaca 28260gtctggctga tccaaacttg ttcggcttta
agatggttgt agaagatgct gtgggtattg 28320ctctgattac ttacaagatc gattgtgaga
aacaagcaga agctatggct gaccacgaag 28380aaattcgtaa ggctgaaatc tcccaccgtt
actttaaagc tcttaagctg gctggagcac 28440tggcatttgt tgaccaaagt tcattcattg
aaatgtctca tcttaaacaa gcaatcttgc 28500ttgtagaaga atccggggca gcattccagg
gtattctcaa tcgtgagaaa gcctatgtga 28560agctggctaa gtatatcgct tctgtaggta
aagaagtgac tcatgctgac ttactggagt 28620cgttgccgtt ctataagagt ggcaatgcag
ctcgtaatga gatgatgact cttgctacag 28680catggggata caaacagcac atcatcatta
agaaaacttt taatgaaggt attgagttct 28740tccgtggaga gactctgaaa gagactgaca
tcaatgagat gatagtggcc tatagtgata 28800gctttgctta tgactacatt ggtgaacgtg
taccgttcga ccagttgcat gtattaaccc 28860aagctcccgg tatgcactgg gtaaaccatc
acatgaagaa cgggcatcgt tccgaagaga 28920acgttattcc aggatttaac atgattgtta
ttgactgtga tggtggagta ccactgcata 28980cgtgccatga actgatgaag gaatataagt
tcatgaccta taccactaaa cggcattctg 29040atgaagagaa ccgcttccga ctgattattc
caatgaacta tgagttacac ctcgacactg 29100aggaatacaa agagtttatg aataacgtta
tgtcttggct accgttcgaa acggatgaat 29160ctgctaacca gcgagccaag aaatggatgt
cctgtgagac tggttcctat cattacaatc 29220ttgaagcaaa tctgttggac gtgcgtgact
ttattcctcg tactagtaag aacgagcagt 29280tccagaacca gatgaaggaa gtacagtcgt
tggataatct ggagcgttgg ttcgctagtc 29340gtattgctac cggtaatcgt aataatcaaa
tgattaagta cgcactggca ttggttgaca 29400gtggttggga ttttgcccaa gtacagcaag
ccgtccactc attcaataag aaactggcta 29460atccattacc agatgatgaa ttgaatgcaa
ccgtaatggt caccgtggct aaacgcttcg 29520ctggcaaata agcaaacagg agtctttctt
tggtttgaag gactcctaaa ttaaatgagg 29580aaaaataatg tccgaagtaa ttcccaatga
tatgaacact cagctaatcc tgattgcagg 29640attctcagcg agtggtaaat cagcatcact
gcgtaacatc aggaaccagg aacgctggct 29700ctatctgaac actgaggcag gtaaacgtct
acctttccgt aacaagttca atacctacaa 29760catcgaagac ccataccaaa tctgggaagc
atttgatgtt gcatctcctg gtggagaaat 29820ggcagatgat gttgatggta tcatcattga
ctcagcaact tttatgatgg atatgctgga 29880atcccagtat gttctgcctt ctgcaaacac
gcaaaaggca tggggggatt ttgcacagtt 29940ctttaagata ctgctgcaac aaaaagtcgt
taagtttggt aagccagtaa tcattactgc 30000tcatgctaaa gacgaactgg atgaagctgc
tggtgtgatg aaaacgttca tcccagtgaa 30060aggctctctg aagaataacg ggcttgaagc
ctacttctct acagtggttt acgcagaacg 30120tgtagacatt aaagaactgg agaagtatgg
aaacaagatg cttgaaatta cggaggaaga 30180acgtgattta ggctataaac atgtattcca
gacccgtcca accaagaagt ctgttggtaa 30240acgacttcgt tctcctatgg gtatgttcga
taagtccgag acttacattg ataacgatgc 30300ccagaaactc ttagaccacc tggctgaata
ctacgcttaa gcgtttgtct ggttgttaat 30360cacttattag gaaaatcata tgtcattgtt
cagtaatctg aaagaaaaaa ccaaaaacgt 30420tgaagctgct aaagactctc tgggtggtgg
tggcttcggt gcaaaagaaa ccgatatcta 30480cactggtact gtaaaagtag cttacgtagg
caaagctgat tctggtgcag actggatgca 30540gttaattatt gaagacctga aaaactctga
tggcgtgcct gctggcgagt tccgtgctca 30600ggtgtacttc acttctggta atgctaaagg
caacaagccg acttacgaga aaaatggtaa 30660agagtacttc ctgcctggct acactgtcat
taacgacatg atgctgatgg ctactggtac 30720tgaactgcct gaagcagact tcgaagagaa
gattgttaaa gtctacgact acgacgctaa 30780agcagaagtt aataaatctg tcatggttcc
agttgacctg gtaggccaga ctgttacctt 30840cgctctggaa aaagttctgg aaaacaagca
ggttaaaggc gacaacggtt atgtagactc 30900tgacgaaact cgtgaagtaa acgagattca
gaaagtgttt cacccggaac tgctggtcac 30960agtcgtcgag gctcaggaag cagagaaggc
tgaaaaagaa ctaaccccag aactggctgt 31020attctatgca gcatggctgg aaaagaacaa
aggcaaaact cgtgacaaga ctaagagttc 31080tgctggtggt aatggtaaag gtggcttgcc
tcctaaacca ggtgcaggtg ctggcacggg 31140tactgctcct gctggtggta aatcactgtt
tggtaaacgt taatgaaaat cccaattgtc 31200ggtgcagaca ttagtctccg caattggggt
ttagctcgtg ggatgctgga cattgagtcc 31260ggcgtcttcg agcaggtcga acttaaactg
gttcaaactg aagttgacca caacaaacaa 31320gttcgaacca actccaaaga tatacaagcc
gctcacgatt tgtttcttgg ttgtgaggaa 31380tggttacggt ctgctaaagc agtattcgta
gaagtaccag taggctctca gtctgctaac 31440ggtatgaaat cctatggcgt atgcgtagga
ttaataggtg catttcgtgc attgggttgt 31500ccaatctttg aagtatcccc aattgaaaac
aaacttgcac tggtcggtga taaaactgca 31560tctaaggaca cgatgattcg tgctgctcat
gccatctatc ctgaagccaa ctggctcaca 31620gataagaagg gcaaacttct gaataagaat
gagcacttag ctgatgcaat cggtgcaatc 31680catgctggtg taaatctccc agctttccaa
aacctcctta atttaataaa ggcgtaacat 31740gcaaatcatt ttgaaccagt ccgaagtaga
agctgctgta caggcttatg tcgatgatca 31800aatcaatctt gctggtgaca tcaatattgt
catcaatgca gacggtacag cttccgttgg 31860tatcaacgaa gaggttcatg aagatactcc
acccgtggga gtagagaaga aaactcgtcg 31920ttctcgtaag aacccacagg aagctaaaca
tcgtccggta gaaccagagc cggaagaagt 31980agttgaagag gtaaaggtag aagaaaccca
gacctctact ggtggtcaga acgagagttc 32040tacgccggaa cctgaagaag aagcagtatc
tgaaccagaa gcacaagaag aagttgtgca 32100ggaagaggct aaggctgaag aaccagcaga
gaaacctgct actaagcctt cactgttcgc 32160tggccttaaa cgtagttaat ctggtaggtg
gctcagaagc tgctgctagg tgtggtagtg 32220tttatagtcc taatgctact actggtcagg
attatagacg tgtcggctcc atacatagct 32280tttatcatca ctgtcattat cctgtggaaa
tgcagtggta aacacggtgg tgacaagccg 32340ccagagtaac aacaacaacc gaaccagtgg
actttaatac tctccgaagt ccattggttc 32400aactgagact aatatgaaca aatttactat
tcactggctc aacggaaagg ttagttcttt 32460catgggtgga gaacccgttg aaggaaagaa
agcatttcac attgaatcag aaggctgcaa 32520aatcttagta ccatatgctt ggtatagaga
gggtgaagta gaagcattaa aaaagagtag 32580ttaatatcgt gaagggttct gttagaatcc
tttgagatag gaactcctat cataaccttt 32640cttaacttaa tctttgccgt atggcactga
cgctaggggt ggccccccat ccgtcaggta 32700aactatttta agaatgggtt atcacagaaa
atgtaaagca acattcgggt taatcaatgg 32760cctctcccta taggggccat ttctgaatga
taggctatat cccaaaggtt agtatctttg 32820ggataaggct tcagatccct tatcaaacta
accgctcaat gttgagcatt ttaatcacat 32880aagcccctat atgggctggg ttaaatgtcg
gatagcatag ctacctacag catcgaattg 32940ttaggtctta catgcaatat agggtataaa
gggaggactc gtcccctccc tcctattata 33000atggtcgagg tagcacgtag gcatgtgcag
agggtcgcta cttctctcgc tggttcgaat 33060ccagcccccg acctactgac atcctatcaa
tgagtacatt aagctaacgt gcggaggttt 33120acactgtagc cggtgtaggt ggagaagacg
gcagggtagc tacctgtgct tagtgtactc 33180tttgatagtt ttcgtaagcg attatgcggt
tttttagaaa cgaaccaata acataaatgc 33240aaacgatgat gttgttctga tggcagccta
ataagccaag cgtcagtcgg gagtgagtcg 33300tcctgattat caaacgacca tggagtgtcc
tcgtccgtgt attagaaacg gggagttaat 33360caatggtgtg tgatagctca aataggtaga
gcgtgaggct acagtgctga atggtttgtg 33420ggttcgattc ccacccatgc ctacagtcca
gacgatatct gagtgactat aaaaacagat 33480ggagccaggt ggaatccctg gcaattaatt
ccggtgtagg tactagtgtc gtgcacggca 33540taagtgttct ggttcgagtc cagacgccgg
aaccaattat cagttgcacg agatggcttg 33600atatgttcaa gtttggacta agtgtgacac
cgctagattg gagagctgct gcggtaagct 33660atagactcca accgggggtt cgaatccctc
actggtaacc aattcaaagt agcattgcat 33720agaacgcgaa aaggtacagc gacgttgcaa
accaacggta ttgtaaggtt cgagtcctta 33780cctatgagca gtgacgactg caacagtgct
actctgaatt gtaagaatta tggtgggtct 33840ggtgaagtag atagggttcg attccctccg
gtagagtaat ctactagcgt gcttggagca 33900cgaataagac ttaatactgg tggttcgatt
ccatccccca ctgcctattt gaggtgtaac 33960agttaaatcc agttgccgga agtgctattc
tagatacgct gccaatgtgt gaacggtaag 34020gctgtacagg actcggactc ccgcctcaaa
ccaaagtcgc caggtactta tggcaaacgg 34080gtagtaacga actagtcatt cgtcaaacgc
ccaccctatt atgagtcagc aagaacgcct 34140tcacttaaac ttaaacaata taagcttaac
gcaataatta tgtaggggta tgtaaggatt 34200gcgaagaagg agccagttaa agtctggcat
gactcactca tttatgccca cttagcttag 34260acggggaaag caaccgacta ataatcggaa
ggtcactggt tcaaatccag tagtgggtac 34320ttattaatag agaacatagt cctgattgca
ttgcactcta aagacaatgc cggtgagaca 34380gtccggcact ctaattattg aaaacatacc
taacggtgtg agtatgagca gactataatg 34440ctcaattaca gttaactatt atcggactat
caatcctgtt tgctggtaca agcaccggtg 34500gtaatagtta actggcctag cctcattagc
tgcgttaaag ctactaagtc cctcttcgga 34560gggacttttt tatttgagta ttgatgaacg
caatcatgct taagtacaaa ttcaaaccac 34620tctatgaggc taataatggc aacagtaagt
aaaaaagcaa tcgaagctaa gattaagagc 34680gtttactatt ttaatggtgc tgatgcagtg
aaatctgcat tcgttgatcc ttctgcactc 34740ccggcagatg atttggctaa cctgggtctg
gtaacctatt gtgttatcat tctggaaaac 34800aattttaaag ttgaaggtgt atctgcttgc
gtagacccaa ctatctatga tgagcagtta 34860ggtcgccagt atgcatatga aaatgcattc
aataagattt gggaattaga gggctaccta 34920ctacgtcagg cactgcatga gaaggaagag
actgctaaag ctttggcttc ctttgcagaa 34980aacaatacct gtgatggtgg tggttgtaca
atttgattca aaggtgtatg gtagttttta 35040ccgaaaggta ctgaactacc cgacaaagta
gtaaggctca tgaagccaaa acaaaacaag 35100ccctcctagt gagggctttc tcatttgagg
tagttatgtc agaagaaatt aaagtccatt 35160tcaccaacta cattggaacc aagtgtgtaa
atggttttcc gatgagcaaa gaaacatact 35220gcaaacttcg tggttgggat gtaccagcag
atgaagaccc gctagaagaa ggctacttag 35280tcgagtatcc agactctaag tccaaccatc
cccaattccg tggctatatt agctggtcac 35340ctaaagctgc attcgaagct gcataccgtg
acgtagaaaa aggttgtacc tttggtcatg 35400cagtagagct tctcaaatca ggctttagaa
tgacccgtaa aggttggaac ggaaaaggaa 35460tgtatatcac cttagtatct ggtgaaaatt
gggccatgga taaacatgaa aataccgttt 35520gtgagaaacg ggattggctg ggtattaaaa
ccgttgataa ccagtttatg ccatgggttc 35580catctcaatc agatgtatta gcagaagatt
gggtgttagc tcagtaaaaa taaaagcccc 35640tcattagagg ggctttttca tttagttagt
aaggttgagc caagggttca ggttgtgggc 35700acgtaagcct tgaccgaaac caaaggagta
accaagattg ccttgtgcag caatactaaa 35760gatgttgtcc tgaataggaa gaccaacgtt
accaaacatg gtaggggttg gagccaacat 35820agccatagca gcatgtactg ggttattacg
aatcatggac actgcaactt ttgcagaacg 35880aatcttaaag ttgtagaacc acatcaaacc
aacactttcc atatacccac ggaatcgacc 35940aggcagacgg tcatagttaa tgaactcttc
cgttacacgc cccagtgctt cttcacgagt 36000cttaccttta cgctgagtca gttcatcata
gatgattgct ttagcaataa agtcagagta 36060ctcaaccgtc ttctgaatac cctggaagag
agcagtatcc ttagtgataa gtgcatagcg 36120acctgcgtta cgaacagact taggcagctt
atcagccagc ttttccatgt attcgtggag 36180tttaccttca gtaatcagga tgtcatcacg
accaatacca gcatctgcaa ttgaagagaa 36240ctcaccagct tccaacagag gccagatact
caaacgttta tggctatcag agatggactg 36300aatctcagcc ttgagtttac gaatctggtt
cgggttagta gctgcacgta attctgcttc 36360tgcatctacc tgacgcagac gagatttcag
gtactggtta atctcagcag tcttctgtgg 36420aatgctctta gcaatgttct taaccggtac
accacgagca accatctgat aaaggttagc 36480caggaagtta acagcaggta cgactactga
cttaaccaca atcagagtct tagcttcttt 36540aactaagttc tgaaccaggt tctcaccacc
cattacatac ttataggcac ggttgccaaa 36600gacacccagc atagctttct tgaaggtatc
cagtgtttct ggtgaccaac gggaattacc 36660agaccaggca tcacctacag aagctgcacg
ataacccaga gcatcgttga gcatgtcacg 36720acgtacccat aattcacccg gaccaaacaa
actttctgct ttctgacggg tttcactgtt 36780catcagctta agtgcatcgg cagttactgg
gtccagttta gaaccaagaa ggttaacgta 36840ctgggactta ttagaagcag acatcttaat
gtcgttctca tacatgctat gcaggttttc 36900aatcagcata tcgttgaatc gctgagcctt
agcttcttcc acctgacgac cacgccatac 36960accgattgca cgagcaagat ggttctcacc
ttcaatgtgc ttcagcatgt tagggtcaat 37020ggattgctca taagcaacca cattaccatt
ggcatcatat acaggcagca gtggttcatt 37080accacgttca ccacgagcca gtgctttagt
gatacggtct acggaaggct tgtcagtgat 37140acgaccagct accatggttc ccatagtaaa
gcccgtaccg agatctacac caccagcagt 37200attacgaacg ttctgtaaga taccttgcga
gaaaggagcc tgtgcctgta ctggtgcaaa 37260gtagtagcta cgtgctggtc cacgattagc
agagctaccc tgataagtac caagacgtac 37320ataggatttc tcaatcaaat cagcaaactg
actatcttca gcaacaatca ggttaacacc 37380ctgcttgttc tcgctaggga tatatccttt
gtactggttc aaggttgcac gactatcaga 37440cttagcttta gccatttcat ctttacgctg
accaaccaga taagaagtag caaagtccat 37500gccttcaatt tctgtctgag ccagtgaaga
taacatctca cggtcagtct tattcattgc 37560ttccagtgca tacaaagtaa tcagtttatc
caactgagct acatctacaa cagaacgtgt 37620agtcttacgt tcacccagta aacgagaaat
tgcagtagca ttacgtagca ggttgttacc 37680aaccgtacct ttaatcatgt actgagccag
ttgcttagat ttacgattaa tcagaggcca 37740gtttcgtcca gcttgtttct gcaaatctgc
ttccagttta ttaacctcac ggtcaacaat 37800cttctggtca gtcagcaagt cacggatttc
atccagagac atagtgtcac gcagaacagc 37860taagtcagtt ttacccatac cagtatgcat
tgctttccac tcttcattag tcagcttacg 37920gctgaactta gatgcgatag tggtaggcaa
gtgttcacgg aactgttgac ggtcagcctg 37980tacctgtgca cgtactgcct taatcaaatc
atatacagaa gcattgccct tagtacgtcc 38040gattatgtca ttaaccaggt catggaaagg
ttgccatact ttaccctggt tcattgcagc 38100cataacacct tcagccacga ttgcaccatt
cttctctgta gcaatagcag ctatcagttg 38160tgcagcatga gcagcacctt taaccaatgg
gttcttagta ttagctgcaa catcacgagc 38220accttccaga gcacgggtag acagtacatc
aatagagtcc accagatact ggttagcacg 38280gtcaatagca ttaccactag gagtagcaac
ggaatcatag aaggactgtg cattgagact 38340tgtctgcatg attgtctgag ccagtgcatc
cataccttcc tgtacgttgg tagctttagt 38400atcaccagct acacgagcat tcagactagc
cattgcagca gtaccaatgt tagtcagcat 38460tgcatcaaca gtattgcctg atttcttatc
tgctttcatg acaggaatgt cagccagtac 38520tttacgtact tcttcactta ccattgccag
accaacaaag gtaggcagta aagaagaacg 38580accctgtgca tcaaactcaa tgttgttagc
acccataatg gtatcgaact tctgctgtgc 38640atagtaacgg tcagcagggt tagtactatc
cgggtcagtc atgaacgctt caacggtcag 38700gttcttagta acatgtgtgt agtattcctg
tgcacgagcc atagcagcag ggttaacagc 38760agcctcagta gccagtgcag caacaatgtt
agtaaagagg cgttgttcct gcatattcat 38820ggtgaagcca tgagcctgaa catcacgagt
tactttagtt gcattcacta ctgcatcaga 38880aaacttacct tttcgaatta cttgctgtac
tggttctgaa ccgatgtagt cagtaatcag 38940cttatcaaaa gtcttaccta actcttccag
acgagtattg tcaccatatg ctttgttatg 39000gaacagggta gtgtctttgg ccacagcagc
agtaggagct tgtccacgca ttactacagc 39060agagttaaac agtaggccag agaacatatc
atctgcatta gccggagctt tcttacgtcc 39120aaataccaga cgcttgattg cttcatatac
agcctgaacc atagctttaa gtgcagtggt 39180tttcttctgc ttaccaatca attcacggtt
ggttaagccc catgccatgt actcattcaa 39240tgcagcagct ttagccattg ctggttcaat
gaagccatta gacaaatgac cattaatggt 39300attgagagca tcagcatatg cttcacgtac
tgccgggggt tcattcttaa catccagagt 39360acggaactgg ttcatcaggt cttcaatgtt
ctggactgct tcattaggag tgccttcata 39420gtgagccagt acagattcaa aagtagaagc
atgaaccaat tcatgaacca gagtctctaa 39480agaaggagta actaaataga tggtcttatc
atcaaagtta gtccagccgt atgcattacc 39540tgcttcagca gcttcaatat cttccggtgc
tgggcgagta atattcttct caatagcgta 39600ggcatccagt tgagaaggcg taccataaac
aaccttgtag tctttagcag caagggattt 39660ctgtacttct ttcaatactg cttgctgttc
tggagacatc tctttagcca gcttagtgat 39720agcagtattg gacaacagac gtacaccaga
cttcagtaca cgaccaacct gttccattgc 39780cggtacttct ttagctggtt ctgctttagc
agcacgtact gcatcacgac gcttatttaa 39840ttcagcatca aacagttcgt tcagctttgc
tacttgctgg tctacagtca gaccttcaag 39900agaaatttta ccgtcgttca catatggagc
accaacagca gccatctggt caacagtgac 39960ttgtacctgg ttcattacct tgtggcgaat
atctacaccc agagcaatgt tacgcaggtt 40020acgttcaatc tgttcagcac caacacgtaa
caggtcatca gtagcacctt cacgctggtc 40080atattccaga gcagacttag cgatagcctg
tttggtttta tcagacagtt tgctgaagtc 40140tacattcttc atgaacttag cgtaggagtc
ataaacattc ttgatagggt taccctgcca 40200ggaagtatat acagcctcat tagctttacg
acttgcatca gtaatgtcat tgataccaat 40260gttcatacca tcaaagattt tcagggtatt
cttcggagca cctttcatag tagaaagggt 40320ctgcatcatc atgccatcac cagtaccgat
ggtcataaat ggaatacctg ctacacctgc 40380ctgtgaagga gcatagatgc tcattggcac
acgcatacgg tcatctaagt tggtagccaa 40440tacttggtta gcaacatcag tattctcact
accagcaatg tagaaagtct gtgaaccagt 40500ctcaatcatt ggagccagtg gagataatga
agcctggatt tcattcagtt ctttctgtgt 40560taagaaatca cccttcttcc atgttgggtc
tttttctttt tctgccagtt tctcctgaac 40620acgttgctgg aacatgtctt gcagaaccag
agattgaatc tgggtagctt tctgcaaatt 40680ctcagtagag tacatcagac cttcacccac
ggtatttcgg ataccagtac gcattggttc 40740aacgaacaga tggagcatat tctcctgcaa
gttcttcaga gcattacctg atacagtaaa 40800cttctgaggg tctaacttac cagtcagagc
accatctacc tgctgaatag acaactcacc 40860tttacgcata accggaacat taccagttag
tgcttccata gaagtaagga aagtatccag 40920catagcctga gcatcagcct cagacgcagc
ctccttacca aacatagcca tagctgggga 40980aatgctaggg tcagcagcac gggcttgtag
aacgtcactg aatcgttcat agatgacatc 41040agtaattgca ctaaccattt tgcctgcaat
accacgagca ccagaaccat agatggtaat 41100agtcagtggg tttttagcga taccacgttt
caggtccaga gtaccatctt cattcaaatt 41160aaagtcttta atgaacaggt ccattaactt
ctggagatgg ttcatctggt tcataacagg 41220aacgttgtta cggtaagtac tacgtaattc
attcagtgct tgttgcagac cattagtgga 41280tgcttcatac aggtcaacgc tgtcctcatg
cgaatgatgc tcgttcatgg tcttaccagc 41340tttaccgaag aacaaaccac ctttggcagt
attcttaatc cagtctgggg tgaacttacc 41400accagtcatt aataccattg cgttgattgg
tccgttggtt acaccatcag cttcaacgta 41460cagtggggta ttaaagttgg tacggtcatc
actgttaagg taacgggcat attccatcag 41520agccataaga gctacgaatg atttatcact
acccaaagaa gtcttcagaa tatccactgc 41580atcggcaggt aagtgtccag acttatcaaa
ttcaaccatc atatcaacgg caggtttaag 41640tttgccttcc agagctttag tcagctcatc
tgacataact tcacgggaca ttttatgaac 41700tttgataccc aatgcctgag ccagtccaag
ctggaagtca gagaacgttt gactgttttc 41760attacttaag tcgatggtag agaaggttgg
aaggattgct tcacgaacca gcttactgga 41820ttgtgggttg ttcttaccaa gcatctgcat
acgacctaca cgggtcatgt tgtagccgta 41880gtgaataggt gtatccagtc cattctcctg
ttccttaatt tgattgatta caccaaacaa 41940ggaatcatac gccatggaca cggacaggtt
tttaccttcc agagacttag cagtattcac 42000attcagcaat tctggattaa gagcaccagc
acccattaac tccagaatgt tgtcacaacc 42060caatgcttca tagaaattaa ccatcggcat
atgtacacga aactcagtag cttgttctgc 42120tttgagagca gctttctgtt ccggggtatt
cttaacagcc gggttacgta actgagtttg 42180agccacggaa ggaatgtcat cacccagata
caatttctct actggttcta aaaggacagt 42240ttcctcaata gcagtaggga atttattaat
ggcatcgtta tcatccagct tctcaatggt 42300atacagacca acagtcttat tagattctgg
gtcaatctca gacacatcta acatggactc 42360tttaacttca cccatctcaa taagggaaga
cagaatctct gtagccattg ccattgggat 42420acccttggta taccccagag gtgcattagg
attacgattc aaaccccaat aggattcaat 42480cttttgagcc agtgagttag tagcttcaac
cagggtctga gcattctcga agtcagccag 42540aataccttct ggtagcagag atgcttccac
accagtaata gcagctacgt ctttcatgtc 42600tttaacagca gcattctgtg tagcagtcaa
tcgccattgc agaccagcaa gtacagcagt 42660ttctaatagc tggtcattga acttaaaggt
gtcaccatct ttctctacga tgttaagtaa 42720cttaccacct acccaacggt tagcctcagt
accttccgca aaacgtttac caacgttctt 42780gttagcaagg aacttagcca gacggtcaga
cagagtagtc ttcagagttt caccaaactc 42840aaataagtcc tgataacgct taatcacatc
acttgtgaga gcgttatttt ctttttgagt 42900gaatgcttca aagcgagcag cagaagacag
tgctttctta acatcagtca aaggagattc 42960agaaccaatg gtacgagact taggctcttc
tggtagggaa aatgctttaa ggaactggtt 43020tggtgtctta tcattattat agacagggaa
cactgtttcc atttccgata aaggtttacc 43080accaacggta tagtctgctt tcagttgagc
cagtgatact tcttccttct caacgttatt 43140ggtattacgt tcagacgtat ctacaccaat
ctgaataagt gcattgtttg cttcaacgaa 43200agattgaaca gtattgaatt gttcagcagt
cagtttctga ttatcaaaca gttgcagaga 43260tacaaatggt tgctcactac ctttatagaa
ggtagagcga atagcaccag tatccagaga 43320catatcacga taaccatcat gcccaaatac
gtcggacaag ctgatatggt cgccattaga 43380accaaccaca gtaccatcag caaacaacca
agggctttct ctacggtcac cattcttatt 43440aagagtttcc tgtgaaatag tattttcttt
cacataagca tctctttcag attcagtagt 43500agagacttct tctgcttttg gactagtctc
tttagtagga gcagtcttat tttctaccac 43560tggtttatct ttaggctgta cagtagtggt
gtcctcttgc actagtgcag gagtttcatc 43620aacctgattc actggttcat tagtctgttg
agactgagtg gcatcaagac caccttgacg 43680gaactcttta actacctggg cagctggctt
gttcagacgt gaatccaacg aggtaacttt 43740cacatgagaa acattaagct ccgggtaagc
cgtagcaagt gcgttagcaa tgtcagctac 43800ggtcttagct tccagtgcta cttgctgggc
aaacttaaca gacttgatat cgtaaggatt 43860gacaccgagg ccagtacgac tacgtaccca
ttcacgggaa ggcgagagag cctgataatg 43920aacagactta ttcttatccg cattccccga
aatcagatgc tcattcaacg ctccgacctt 43980attctgcatg tgctgggcga acttcataaa
atcgctcaga taagcggagg ccaaatcgaa 44040gttaccagag ttatacgcag aacgaatgcg
tttcgcatgt tgcaacgcag agtactgtcc 44100ttcattagaa cgagactcat cggttttaat
ctgtttacta acaatatctt gaggacgtaa 44160gcctagttct tctgctttag cgtcgaactc
tcgtgcaccc tgtaataagg cagcagcaga 44220ttgcagggca gcacgttgac gattccccag
tgtaatctta ccttcacttg catgtttcag 44280aaccatgttc actgaatctg catccagcac
ttctggagat acatcagcag ccatagcaat 44340gttgtttgct tgggcttgat tagcttcttc
agtagctttc agcttacccg cttcagcctg 44400ttcctgaatc atggaatgga tggcacgaaa
tgcacggagt actttagggg tattctgtac 44460gttagccatc aggccggaga attggtctac
gattgcagca gcaggggaat cttgttccag 44520attagccaaa gcacccgggt cacggttaat
gaagctatcc atagacatga tattgtcata 44580catctggatt gcagcttcca tttgaacgtt
agggtcttcg gcagtattaa ccaagtcagc 44640catcttctga atagcttcta cacggttagt
ggaaccagaa acagcttcac ggattgctgg 44700gttagtttgt tccaattcta ccgggtcaaa
cttcattgcc tgagttaagt cagcagcata 44760ctgggtagca gcagctttct cttctggaga
aatatccatt gcatccactg cttcctgaac 44820agttgcttga gcttgttctg cctgagcagt
agcttcttgt gcagcagcgt taaccgtagc 44880atcagatacg ggggatgctt gttcattacg
cttagcaact tcttccccac gttgaaccag 44940gatgttagta ataggagaag ctacttttgc
tagtgcctta ccagccaaag aagcaccagc 45000cagggtagta cgtacagcag gaccaacggc
agcaccagca gccttaacag ttgcaccagg 45060agcttgagca actccagcag aaccgaagcc
atacaatgaa ccaagaccgg tttgttcacc 45120cacaccttta agcaaatcac gcttagcatc
tacattacct tgaattgcct tgttctgtgc 45180aaactgactt gtaccagact ggataccttc
ttctacagtc tcacgcagca tgttagaacc 45240agcaccagcc agtgaaccta ccttaagtgg
attaagttca aacttggata ccagtggacc 45300agtaagagca gcaaccggag cagttaagcc
agcagcagta atacccgttt cagatgcagt 45360ctggcgacgt gcttcttcag gagataaacc
gtctttaata tgctgttggt atacaggaga 45420tttagcagcc aattcactga atggcatttc
cataatttca ttagcagtct gttggtatgc 45480accaccagct tccataccac caattgctat
agcaggtgca gcaacacgac cagtagctaa 45540tgcagtacgt gcaggacgtg aacctaactc
agcagctaag gtaatgcctt taacagcttt 45600atcaccacct accattactt taccgagtgc
agatacacca cgaatcaatg gaccgccggt 45660aaagagagaa ccaacaccct cagccagacc
atcagtagca gccatgccat tggacaaggt 45720gttagcaaca gaatcgtaag catctcgacc
aatgcgagac agggaagcaa cgagatcact 45780ttctccttta gcaatatcct gcttatagag
tttctcgttt tcctgtgcag atattacgtt 45840ctggttctgt actacattac gacgagcatt
taatgcatca gactgtaggt tgtgtacacc 45900tttattcaac cagtcaagac cggaagcaat
ggttgccccg gcattatcat tgaccaagcc 45960tgtaccaaga gcagcaatgc ccccaagagt
attagcaaca ccaagaccaa caccagaaag 46020ggtatcacca agtgcttcac catatgtacg
actcttagtt aaatcacgac gaacagcatt 46080agctgcattg acacgagcat tgagaatgtc
cattccttgc tcattgccat acttgtttat 46140gatttcaaga ggagaggcat tagtaaaatc
tgcctgaaaa gaacctgggt caaatgcacc 46200agcacctaag ttaccagcac ggcctgcctg
taattggtaa gcgttctgtg gggttagttg 46260gaaggggctt tgcccttgtt cagcttgttt
ccgagaggtt gcagtagaga catcgacttg 46320cttagcagta gtgatgctgt ctgcgaaacc
tgccagacgg tcaaatgttg acatagcatg 46380attcctgttc aaacttatat cgtataagaa
gattcactgt acagtaagtg aatgtaagag 46440aaaagccccg gatggggctt tattagtatg
taagatttta tcgaggttgc atatagggag 46500gtaaaccctc tgctaatttt cgataacgtt
tagctcgttc ttcatcttca cgactacgtt 46560cacgttctac tgaagcatta gaacggaagt
tctgtggaac cagatttccc tggtcagcta 46620ccacagtatt aacagcagca tctaatgcag
ctcgtgccct tgctaactca gcctcacgag 46680cagggatagt agcagcaagc ccgctttgcc
cagcagctac acgacgacga gtatcagcaa 46740gtaaggcttc tgcttgtgtt acctggttct
gagcattagc aatattaccc tgtacctggg 46800taagaacatt atcacgagca gtagcttcaa
tgccttcccc acgacgtact gaacgtgcta 46860gttctctggc aacggtatca tcaatacgaa
tacctccacc accttcattg gagatgaacg 46920gattaagtgc atcaagtgct cgactaatca
taccttccgg tacgttggta gtagactgct 46980taagaatctc agcagctaca gcaggagact
gcccggattc ttgcataaca tcattgagtc 47040gagcaatcat ccatcctttt tcagaacctt
taaaagcatc cttaagaaga cggtctgcta 47100cttcaccaat agtagagttg tcacgtaatg
cacgagcata attagcagta atggttccag 47160tgttatcttc agactgacgt gcagcaactg
tattgcctgc catgtttgag actacttgct 47220gagtctgtaa ctgatcaagg ggtgctccta
cttctgcctg tgcaattaac tggcgtgctt 47280cactccacgg catatcctta taaccaccta
cacctaagtc aggcaatgca gcccaggtct 47340tagaaaggtt accattctta cggtcattga
agattgcttc agctattttg tcctgtactt 47400caggagtcat ttgctgatta cgccaatctg
aaccaagcac tttaggggca tagtcttcca 47460gagtagcttt attaatctga aatgcaccta
caggagaatg accttgtgac gggttattaa 47520tcatactctg ttgatgatta gtaacctcac
ctagtgtcat ctcagtaata ggcttatcag 47580tagcagcaaa ggtataggtt gcatcataag
gagaaccaga acgagtacca gcagttcccg 47640gagcagagcc agaaggagac gtagcgtaag
tacctgggaa tctctggttt acacctcgca 47700tgattgcagc acgtacacct ggagaagcat
tctgcatagc agactctgcc aatgtaagag 47760catccatagg agtggaagca ctacggaaaa
tatccgtcag tataccggat gcagcttgtg 47820aatcagcatc attacgttga gccacaccca
aatcaaagcg gttctgtgct tggttaatag 47880cctgaccacc ttgcccttga agacggctca
gcatatccag ttgttggtcg gcagggagtg 47940cagataatgc atcacgggtt tggtccaatg
cagtattaat acggttctgg tcaccagact 48000gataagcctc agacaataaa cgaatagctg
gagaagcatt atctaatgca gagtcagtat 48060tctgtaaacg accaaaacga taagcattgt
agtcattaat tccctgctga ccttgttgag 48120tcagaagagt acttgctcgc tcatccagat
tctgcagggt atgttggttt acgagtgaag 48180ggtcaacccc ctggaacaat gcaccggatg
ccagagcatt acgatactca gtagggtctt 48240gatactgcat agcattcatc atgacggcat
tgccagcctc ctgcttggca gcgttctgga 48300agttacccag tgcatcactt aagccggagg
tggcgttacc aatcatgttg ccaaatgtgc 48360gaatgctgtc accaactcca gagaagttag
gtgcatcaac attacgccat gtaatttgag 48420ccatgatggt ttcctattaa cgagttagct
tattagctgc aatgtaagca tcagctgaag 48480actggtcacg gttctctgca acagcacgac
tacgagcacg gtcttccagt gcagtgttat 48540aagacttaat ctggttgttc aggttagtgt
tagtaacact cttagcaaag ttcaactggt 48600ctttggctaa cttattggcc tggaaaccac
catagatatt agctaatgaa ccaatagcac 48660ctagtcctaa ctggaaggtt ggtacgttca
gacccaactg attagctgaa ccagacagga 48720aagaagtagg ggtagatgca cccaaattag
tacccatacc aatagctgca cctgggttat 48780agttcatagc tggggtatta aagttcggat
tgttattaga catccaagac atagcagctg 48840gctgtggggt ttgattgcct gttaagaatg
acaaatccat ggggattctc ctgttaaaca 48900aggtcagtat taagggtcat gtcagagaag
ctgccaacca tgtttaaaga catgtcagct 48960atatctgaac cagtcataag agtacgagat
aagaaagaat ccatcgactc cattgacacg 49020aattgcattg ggtcaattac accctgccca
gcagtaccaa acatttcttc atactgctta 49080ttgattgcca tcatatcagt attgtactgc
tgcattacac tctctgcttt ctgaatagta 49140gcagccgtag atgcattaat atactggcta
ataccattac ctactgaact ggtaagctgc 49200atgatgttct gagcattcat catttcactg
gctaaggtag ataaggatga accagtagat 49260agagcagtac caacattcat agctaccatt
gaagcaacag cagcaatgat aaaacctagc 49320ttatcaccaa agagtgaagt ggatacttta
gtgataatag ataccagaat cattgcagca 49380atggcgtttg ctacagcacc tactattaca
gcagctaatc caacaaaacc gagtgacgcc 49440ccaactgctc catatgcccc aagtatacct
gcaccaccag tacccatagt gaatacggat 49500acaacgacag caaccacaac aaccacaatc
ttaaaggcag atgtttgata ccacttctgc 49560ttaaccttct tatatgagtt catcactaag
taggagcagg cagtggatag ctgagtactt 49620cgaatcagtg acattgaacg atagatgtta
gtatgtagtg gaataatgaa cccactctcc 49680tctgcatcac ccattgcttc ggcaacatca
atatgcactg acttattctt atataccctg 49740ttattgtgat ttagacccag tactctaagt
ttacggtaag tgttattacc atcctgccaa 49800agcagttcat actcctgcat agaatagaat
gtagtggtaa cctctaactt cttagctgaa 49860ccggagttac tagcagttcg tatgttcttg
cgagtaagag ttatatcccc tgcatacctg 49920gctcgaagtt gtccttgctt agcaccagac
catgcttgcc ctgagtgggt agtttcagat 49980acgtaattcc aaccaatggt catgtcatac
ttgtaatgct tattactatg taccctaaat 50040tctctcttag gtatgactgg gtattctggc
agtggtggag gagttccaat agtgtgccct 50100tcagtcctat tccaccattc tacatatgca
tcaactgcat cattagctgc ctgataacct 50160gcaatcactg cctccaaggt tggataggtt
gggtcaggtg ggaatgcttc agtagccatc 50220tgaaagaaac ggtagatgta ttccttagca
gtatcttcag gagtattaag agatacgcca 50280aatgtgccat aaatgtactg aatatcacct
atatcatcat tcttcttaag ctctgttact 50340accttatcaa tttttccacc ggtagcttta
taaagagctt tcttacagta tggatagatt 50400gggtcattct ctacccattg tttatcgttc
ctgattggaa taaatggata gaatcgatta 50460tcggtagcct cagtatcgaa cagtgagtct
agtgcaacgt taccagagtt ctgcttatag 50520ataagcatct taggtgtacc aacagcagtc
atctctgtat ctgtacgagt agtctgggag 50580gaatacttac gaaccagtgt ctctgtggtt
gtagtaattg tatcagtacg agtaacacca 50640ccacctatat caactacgtt ggtagttaca
gtagtgctgg attgaatctc accaaacttt 50700ttatgagtca ttaccctgtc agatacaact
acagttacat catcgggttt ctcagtgaat 50760cctttacgat aaaccttaac gtaagagttc
cagttatcca cacttggttc agtgacagta 50820ttgtcttcat ctggtctacc atcagagtaa
acagagtgag tatgtatggt cttagtaagg 50880ttagttgtat tagctgttgt atcacttgat
atggtagtcc atagcaaagt ggaaggtaag 50940tcaccctcac tatcatagac agtagtcggt
cctactaccg gtggattggt agagggagat 51000ttatagaaag agtaatcagc atagagatac
aaagcacctg gttcaaagtt agtaggtgtg 51060aatttaatag tagaaccacc atccagggag
gtcatagtaa tttcattagt atcttcattg 51120atgtctatgt cgaatcgttc catgattcga
ctaggagcat tttcataaag atattggtca 51180caccattgct caaagtcagc aaagccaatt
tctgctgcct gaacatagac agactctcca 51240gcaggtggtg taatttgacc ctctataaca
gtagggtcaa tcttagccaa tacacccaac 51300gaagaaccag ccataccaac ttctgaatca
tagtggttct tactccagct ggagaacaat 51360cgcatacgaa taccaggtcc attcagataa
ctatcagaga tagtgtctgc catagtgaat 51420cctgtattgg aaacaatgtt accgatgact
acagtcttca tatagttagg acgtttgtgt 51480atatcccctg ccatgttata gacagaggat
gctacgtata ctttagtctt cccactgaat 51540aagcccatat tagttcagcc cgttgttagt
cttcagcttg gtcaaaatgg tatcaatact 51600tgcgttagtg aaaccattag gaggattcaa
gccttcatca atagtcttct gtgtaatcca 51660tgcatcagta aacaacttag atgctttgac
ttctgcatca cgttggtaag aagtaatctg 51720ttgagagtac aactctttct gtttacctac
agaaccagta acagtagcac catcactacg 51780agtatccagt gtctgtgcac gttgtgcttc
tgtctgctca gtaagcaatt taagctgttg 51840aggtaacatc tgattagcat taaacaatgc
agcacaatac gtctcagact cagtagcaat 51900cttcatctta gtaagagcgt attcactctt
agcagacaat gcctgaatct tagccagtac 51960aaactgagcc ttagaagtag ccagttgaac
acgagcagta actgcctgaa tctgtgccat 52020agcagcagcc cagtatgcct ggtcacgtcc
aagtaagaac tgaacagcat tactcatgca 52080tgattccatc atagcaatgt atgctttggt
atattcacca ccagtaatac ggttagcttt 52140aaactcagct ttaaggtgat tgtgagcaga
ttccattaat gcatcaaacg taccactgcc 52200tcctacttca cgggtagtaa gagattcatt
ggttatttta gtaatagccc caaagattgg 52260agaatcatct ccaccaggga tatcccattc
aggaccagac atatcaatat caggaagggt 52320aaagtcatca cccttagtta actcttctag
gagtcggtta gcttctacct cagcagaaca 52380agacataatc attcctcttg gttcaaaaag
aaacggccca cggagttacc ccagtgagcc 52440gtgttgaact gtagcttata cgttaatcgt
taaggctacc agcagcaatc tgtgcttgag 52500ccagctgagc caattcagct tcagtcaacg
gaggcagtac ttcaatggag aactcacgtg 52560cttctgttgc acgaatgtct ggtaagccat
tcttaccttt acgagtagta atgttaatga 52620acttacgttc tttaaggaac tcgtagatac
agtacggaat atgataacca ttgtcggtta 52680cttcaccgaa cggaacaaac ttacgtacag
tacccatata ttcgttagct acggtgataa 52740tttcacccgg caggtctttc ttcttagggt
caaggttctg gatacgtaca cgaatcagtc 52800gagtctgttc tgcacgaatc ttctgaccca
gggtcatctt cttaacacca gcttcctgct 52860tagcacccaa tgggttaaca gcagcttctt
ctactactgg ttcatctttt acttgtgcag 52920cttcaatctt ctcacgaagc ttttcaacgg
aaatgttgtt agagaactta atattcatca 52980acgttgcacg ttgcttaaga acttcaagtt
cgctaggcat tgcaatatcg ttaacggtat 53040cttcgttgcc ctgtacgttc acttctacgt
cagtggtcgg tttatcgtta atgctcatgt 53100tcatatttcc tgtggttcat ttagttttat
taagaggggg acttatgtcc cccttgtttt 53160atttcggact attacagagg agcaacagtc
ttaatcagag ccagacgttc tggacgttta 53220accaggatac catagtacca cttgatagaa
ctgaagccag tttcgccata cgggtcatta 53280cggtcagcag tttctttacc cggcatctta
gtcatgatgg tgaacttaac agacttacca 53340tcagtctgga agccaatggt agagaaggag
tcatcaccaa ctaccagcat cgggaatacg 53400tcgtagtgtt cctgaccaga taccatagag
gtacggtaac ccgggttagt agtaacctga 53460gcaccagcac ctgcccaatg cagcatctct
ggaacctgga tgatacggaa cttatcaata 53520caaccaattt caccattcat cagagtacca
gcatcagcat agtgctgaac ttcgatgaat 53580gctttattac caaacaggtc tttcatcgct
ttcagttctg gaaccagttc agaaccaacg 53640tacattacac gagtaccacc gagtactttg
gtatctgtca gtttagaacc agtgatgata 53700gtagtctggg tcggggtacg gttctcagta
agaatctggt caagacgcat caggttctta 53760taagaaacta cagacggggt agaaccttca
ccagtaatgg tagcatcaga gacggcagca 53820cctgcataca gtacagtacc agcagcagcc
agcaggtctt tctggagaac agcttcagtc 53880aactgtacag caccgttcat cagttcacga
gacaggtgtt ctttcagttg gtcatcagaa 53940tcaaagtcca aggactcttg agtaaattcg
tagaagaagc cgaatttatg aatagagcct 54000tcacgagcca gacgagtaaa gcctacacgg
ttaacacgac caccattctc tgacagcaga 54060ggcagtttag aagtgatgtt accaacgtct
ttggaagagc cataaaggtt accgttaacg 54120atggtagcac cattagcatc aataccctgg
tcgttaatgt tcttatcgtc gagcaaagga 54180acatactcgt acaccttaac agtcttaccg
tagttcttcg gcatgttaac ggtgttagcc 54240agaggcataa aatactggtc tttacgagac
tgaataatag ctttcttcag ccagtaatag 54300gtattcatct ggtcggaacc agcaccatca
atgctagatt tctgaccgtc aattggagcg 54360ttatagttta acatatcatc tcattcctgt
ttaaagacta cccggtactg ggagtttagc 54420gaaatcttca tcactcatag cgagtgggtt
tacaataggt gttgctttac gtgtagcagc 54480ccgattaagg gaagctgctt tagcttgctc
actgttagcc agagtctgct taggttgtgc 54540cacacgcact accggctgaa ctgctggttt
aactgcttgt actggttcag gcttagctac 54600ttggttaaat gctccttgtt gtgcaagaag
gttaccaacg tagttataag cctgaataaa 54660cggagtacct accggaatct gacctaacac
ctgaagacga ttcacttcat tagcaatggt 54720gtcataaata ccattctcac gctgttcgtg
aatggtgtgg agtaagccac gattctgata 54780aagagcatct ttactggcag catcccatgt
cgagctaata acacctaatg tagcttgtcc 54840ttcttgagta gactttaagt cgtcaatttc
ggttgcaaaa tctgcttcgg tgtctgtaac 54900acggtgtttg ccaccctggt agttaatttc
ctcttctgga ttaaagtcca gaggatctgt 54960accagagtct ttcaacaact ttttaatggc
ttcaggattc ttcttatcca ggtcaatcag 55020gaaagaaagt ttttcctcat ccattaaacc
gttgttctgt agcatcagca ttaccttacg 55080gtacggctga agttcttgca tcttacgagt
atagttagca cccatctgca tcaggctaat 55140ggcctcctcc ggtgaacggg gagtaatcat
tttgccgtta gctttaaaag gagccatcaa 55200cttctcgtaa ccttccttat agttgaagtc
agcaggcaga ccttcagact gtttgccttc 55260ttctttctgt tcctggcctg gttcagcagt
agaaggttca gcttcagtaa tcggcttacc 55320gttactatca acttctgtgt caacaacttt
atcatcaact ttatcagaag ttaaagaatt 55380tgctgaatca acttcatctg gttcagtttc
aggggaggtt tcttctgttg gtaggtcttc 55440aacagcaggg gtgtcaactt cttcatcagg
agtctgtacg ccgttggttt ctgggttgtt 55500ctgagtggaa gtatcttcct cagcaataat
agctggggct tccatattca gaatctcatc 55560atccgacatt gcgagaatgt cggaagctgt
ggttgcagct tccgtagtca taggaaatat 55620ctccggttaa ttattcgtct tcgggttcag
cacgaactgc atcgagttct tcttctacct 55680gaagaataac gtcagcttca ttttcaccca
tacgaatggc gaggtcgagc caacgacgta 55740agtgaccagc agcttgagcc atgttaagtg
catctgcacg gttattcggt tcaagcagtg 55800ggtcaccaga ctcctgcacg taacgtgcac
aatcttctac acagaactgt cgaaggatta 55860ctttacggaa cagtggattc tccagaagtt
tacgtacatc ctctgcgtgt gcaactgcac 55920ctttagcagc ttccagtcga tgttccagtc
ctgcgattgt tgattcttta ctcatgtgaa 55980ggtctgcctt ataagttcat accaagagcc
gatgctgggt cttggctagg gtcataaaat 56040tgggaactaa gagaataagt cgggtcttgc
tgtgcagcta agtcacgttc ctgtaaagag 56100ttcccgttag tcaaagcgtt atatccgaca
gcagcagata tgttgggggt agtttcacct 56160tctttagtag gtgttgtcaa cgccttagtt
atttgaaggt tctggtttcc ttgagattgt 56220gctttttgtt tttccatatc acgagcatgt
ttagtaccag attcctgttc cagataatcc 56280aggtctttaa ggtcaccact agaaatagct
tctttagcct tagcattatt gagtgcaatc 56340ttacttttca actcttcatt ctcaagctgt
gctttctgaa ttgctaattg cttaagctgt 56400tcttccatag ggtctggttg tggtcgccag
gtacgtaatt catgagcaag gtctggcata 56460cgtttaagtt ctgcaatctt agctacaaga
gataatgtaa tagtctggtc aactgtatta 56520cctaacgttt gaaccatgaa gctcaagtcc
tgagatttct ggttatcaat ttcagcagta 56580ttaatatcaa cctcaatatc gaaattacct
ttaaggtctt cacggttaat ttctacgtac 56640tgttcattgg taatacgtac tacttccttc
tcagataaga atacggcatt cattgcacaa 56700atcttagtac caatgtctgc cataccctta
gctaatcgac gaaggattgc catctcacgt 56760ttggatgctg catcgagtgc accacgaata
ccagcagcca catctccata agctgcacca 56820gttacacctc cagagaatgc tttaacacca
gttagtgctt ctgcttcctg gttctgcatc 56880tgagtcataa ctattgccga ctgaggtaac
tcagggaact tgtgttccat aatggcctga 56940ctaggattgc cctgcattgg gttgtattca
tagtcttgcc catcatcata tcgacgacgg 57000tttagagtat ctaacatccc tttaggataa
ccacgttgcc cgtttgcact tcgacccaac 57060aggtcaatca tcccacgcat ggttgcacca
agaattgctt ggttatcccc caacagttca 57120gcatcagctt caccgaagag ttcacgttta
cgtggcatat aaggaacaac taccaaaggc 57180aacttaccat ccgggaatgg gttcttttcc
atacgaataa gagtagaacc aatccaggta 57240gcaacaatag gctccagtga accatcatca
ttgatgtcgt agaatcccca gtactcataa 57300gcaactactt tcttacgtag tgcatcctta
aactggaagt caccaggagt tttactttca 57360tggtctgggt cagtcatagg actggaactt
tcccagtcaa tcttatccaa attgtgataa 57420cggtctttgt tcttcatgag gtctgctttg
cacgtctcaa acgagataac tgcatacaaa 57480gccttgtcca aatcaccatt acaactcggg
tcaataacta cgttattagg attaagcatt 57540tcaacagtag gtctgttaac cagtgccttc
tctacctcta cctcagtaac tccagtctgg 57600attgcataag tagcttctcc agtttcattg
aagtagttaa cagcttcctt aatatcttcc 57660ggcattgttt cgtcatactc acggggattt
tctgcctgaa gctgtaaagc ctgctgaaga 57720atatctgctt gttcctggtt ctcaattgga
tacaactgga agactggtgt ttctgtttta 57780atcttaacgg tcttacgttc ccaaccaata
cgggcaatac cagtaccatc atctacgaca 57840ctatgtacgt aatcatccac cagttttact
ttattaagct gggtacggaa ttggtagtta 57900agaactaatt cattctgtcg tgcagctaac
tcatcctcaa aagtaacagg ggttacctta 57960aagagtttat tagatgagag aaatggttca
gataatggtg cataacgcca ctctgcctga 58020cggcgaacca gtctaggttg gacttgtgag
cgtcctttaa cctttggggg tttagccttg 58080cctttgactt ccatcaagtc attccactca
cgaatctgag ccatgattgc atcgtgagca 58140ggtttagctg attccaaatc acccttcagt
aattggatac ttggttcctt cttccagtcc 58200gttaacttct ctgattgagc cgggtctggt
aaaggcttaa aagtgtcttg gtgttccata 58260gttattcctg ttcaaaaagt ttacggtcag
cctgaatctg tttacccaac tcaattagtt 58320ggctgtcacg gagtctaaca gttgcccgga
gttcttcaac caaacgtctg ccttcttcaa 58380gactgttgtc gagtctggct gcatggcttg
caagacttct gcactcaaag gttccggctt 58440cggcttgacg tttatatacc gatgctcgtc
tttcagactg ttgcatccgg ctgtcataat 58500cactgctaac gcgagcaagc tcgcttgcgt
aattactctc agccgtctgc aaccgggaag 58560tgagtaaccc gacttcgtaa ctatggtttc
tttggagagc attgtatttg tcctgtaatt 58620tttgtagtgc cttctgatct tcaacctttt
gggcatccca cttcttttga acagtagact 58680gtccattgga attaccccaa aagtaaatgg
tggccccgag gaccaccacc agaagataag 58740gccaaccttt agagattagc atcttcatta
catgcctcca gtggcacatt gcctattcgg 58800ttagctaccc aaccatacgt aaagtctggc
atgtttaatg aggtgtagtg gttaagttgc 58860ttagcatcaa gtaacttaat cattacctga
catgctgcaa ctttgcctcg ttttttctgt 58920aatgctttat atgcattaac agtacttgta
ccgactttac catcaacctg tattttaggg 58980tagtctttgc catcacggga catttcatta
agagattgct gcaaccactt agccggacgg 59040gttacacccg tattaacacc agcatctacc
agcttatgtg ttacagcagg agatatatca 59100gcgaaggcta cgaagttagg cttaagtacg
taatcatcaa tgtatatctc agcagccatc 59160tctttagata aatccttcat agaaccatcc
cagccatact cagtagccag aacttctttg 59220tgagatttag ctacagcttg ggtaatacca
tgattggttt caccacctgg gtcacgagga 59280ttatttacat atcccccttc catataaaac
actgccccca ggatagcagc gacaactcct 59340cccactgcac cacctttcgt agcaagtttc
tgtttagctt tcatgtgaat cttcctttga 59400tcggaatcgt attaatctgc ctacaatatt
taaggcaaac agcgcgattg caatattgga 59460accatgggga atatcttcca ggatgtgacg
aggtaatcct gtcagcattg gttgaatgat 59520atctatggta gaaaacataa ttaacccgag
cgtactaatc tggatggatg cccacttcca 59580gcatcttttc cagttaggta ctaactctac
ttttcttttt agcctgcgaa ccatgcgaat 59640atctcccgtc ttgcagcagc caatacacca
ataattgcac ctgccccagc ccacacccac 59700ttaccaaaaa ttccagcacc aacaacttta
tgcttaatgg tgatgaactc ttcaatagta 59760ggttcattct tagctaagct atcctccaca
ttttttagtc tattgcctat gtcattgatt 59820gaatcccgta gttctaccag agtttcctcc
agcttctcac gatcctgcct gtcccttgtc 59880tggttctcaa acagagtctt caatcgttcc
tccagtctga ctagaagcag ttcacctgat 59940tcattcataa aattaccgta gtgatgtagt
tagattcccc aacagggcca tatcataaga 60000tgactcttag ataatactta gtatgatggg
atacatccag acatagtgtc tggtatcaac 60060aacgaggtaa ctatgtctat taaagatatt
ttccaaagtg gtaaggatgt tgtgtgcaaa 60120atgaa
6012526122DNAArtificial Sequencea part of
DNA of novel bacteriophage CJ23 Contig00002 2taagcattac caagcaatgc
atctaccatt gggtcattag aactgttctg acagtaagga 60ttagaccaat ccttctttgc
atcaaagggg ttgtgtgcgt tcatcgccat cctcgttgtt 120caaatttagt ttcggtaatt
gaaatacttc cattagccag atcgtaatca acaacctcac 180cacagatgct ttcatacgtt
tgcagatact ctgctgcctt agcgttagct tccggagtat 240tcaaaccggt gtggtaacga
taacccaccc agttatgaag tgccgtcatt aacgtatctg 300ctaagtcaac ttcctgttct
tcatcacccg taagtactgg gtgcttagct tgataagtta 360cgttaagtgc ctcaaagtgt
cgaggtcgca tacactgaat agtatcaggc cgaggggtga 420agattgcatg agggtctgaa
tcgtcgttca gtctacgtcg attccctgag ttatcaaaca 480catttaagat tttaataaca
tcatcctgga aaggtttcat gaacccatcc atgatatacg 540ggtattgaat ctcaagagta
ggcttagtaa accgggagta agcatatcga gattgaagag 600ggtagtcagt acgaccttcc
ttcatctcca caatgcaact attagttcta agagggaaac 660gactatgcag tcgtaccagt
ccctcattaa tagcagccaa tatctctggc ttactatctg 720gttcaatatc taatctgtca
tcagtgactg caccagtacc cttcaaggta cttaatgcca 780agccacgata tacttccgat
aatttctgca tggttcctca cacaatgtat gaacgtagtg 840ggttaacact atcttcttct
tcgtcatccc acattggatc acccttgtta tccaccatag 900tcatacctgc ttgtggtttc
caaggattga gataacccag catggagata gtatcaatac 960agtcatcctt acctttaatc
ccattaatgg tagctagttt aatctggccc atgaacagac 1020ccataatagt tgaatccctc
aactcttccg ggaagtacat cttaccagct ttgaaccaag 1080gaactaccag gttaaagcgt
gacagcttag aggttacagg acgtatgcct ggcttaccac 1140cctcagagga tgcaaagtta
aagaagacat tacgattaat catttccttc tgaaggagtg 1200aaatgaatcc attctgctgt
cctgtgattt cgactccaac gttttgtggt tggtattcct 1260gaaccagacg gaacaggtca
tcaaagtttt tatccataag ctgacgatta gccacaccat 1320caacccagaa ccaatctcca
ttagaactat aagcccaaac tgatatgaca ctgtagtcac 1380tggtctgttt ctccgaagta
gcaaagtcgg ttgtaatata atagttgtag caggacttca 1440tccttaatag ttgctgtctg
ctataccatc taatttcact atcctgaacc agtctctcat 1500cttcagaact aattcgaagc
ataagttcct gatagaagcc tgccaactta ccagtcttaa 1560ctgccatgtc atattgagcc
ttgatgtagt cataagagaa acggtcatcc catgcaccct 1620gaaattcttc tctactgcaa
gggaacttct cacacacagg ccatacgttg acatcccatg 1680caccagactc tactgcttca
atgataatat cttctttatt aaagggagta ccattgaaga 1740ttactttacg acgggtagga
tcaagagcat ggttcacacc tttatagacg gtatccttaa 1800tagcttccat actcgtcttg
gagttagcat caccatcact aatcaagtca tccagtacgc 1860ataatgttgg acgtttacca
tatatcttcg taccacgaag acctgtctta gcaccaaaca 1920gcttaacacc tagacgatgt
ccttctacat tacgaaactc taacaggtta tccgtaaagg 1980tagcttcagg tatccattgc
tgaaggaact cactattctt gtaacgaaac tcaatgttct 2040tacgtgcaga tttagcaccg
ttatccattg agtcagatac ataaatcatt ccttctactt 2100tacccaggct tggtaaatgc
ccgaacactg ccaggaacaa tgtaaagtat tccatgaata 2160cagcagtctt acctgcacca
cggaagcaca gattaactac atactggttc gggctaatca 2220tcttatccaa cattttcagg
tgaactggag gtgttttgtt ggattcaccc tctttaccat 2280taaccaactt aataaagttg
gcaaaggtaa gagcaaactg actaggaaca tagttggaag 2340aattaaggtg ggagtaatcc
acctggtcta gccattcatc cagttcctgc ttaattaact 2400cagacatctg taatgtcctc
atctgcacgt actagtttag aaccagcaac ctctttggta 2460ggtacaccac tattgattgc
attaatttgc tgctcagcta atgcagcaag tgttgccttg 2520aggtcagtta atccagagtt
ctctctcaag tccagattga tattcgttac ctggtctttt 2580ggtttagcta agtgggtaag
gatagagtta gctgcatcac atcttacctt ctcacttgct 2640gctgtagtca tcagttcaac
ctgaacattg attgccttct gataattatc ctggttcaca 2700atccacactg gaaccaaact
ttgttccatg atgaggttaa ctaacttacc cctgtgataa 2760gcagatacat aagcactaat
atctttctcg ctggttcctc gtgcaacaag ttctgcttgt 2820cggttaggga atgtcttgaa
gtaggcttcc ttattggaat aacccatgtg tttataagtc 2880acatactgaa ctgcattcat
gtagtcctgt gtcttaaact taccttcctt cattacacca 2940gagtaggaga taaagttttc
acggaaggac tcagcaacca gttggtcttg ggttatgttg 3000ttgatcgtgt ctaccaactc
ctgggtcaca ctgtttttga agttagcagg taaggcatta 3060acaatctgct gcttagttag
ttcactcata cttatctcgc tacaaagtta tgtctattaa 3120agaagattct tttcttgggg
tgagttaccc cttgaagaaa cgataccata cagttacact 3180ttagtcatga tatagaaaaa
gttcatgata ctaaatagta tgtaatgttc cgatgaccct 3240attcccaaaa ggagtaacct
ttgagaatct atatgaaagc cccagactct agtgacaagt 3300acttcgaact agaccacact
aagattggct tcctttcaga tgatattatc attacacatg 3360atgaagacac ctaccatgaa
ttagctggga tgatgctgtt agagaatcgt ccaggcatta 3420aagtcttaga accaagagaa
acttactctt tagataaaac aatttcatct cttgtgttct 3480tcacaaaatt acctcatagt
gtctccgtca ggcgaagcaa agacagtgtt cggatgacgg 3540tgacgatgtg attagtcatg
ccgcctagta gacagtggcg gtactcgtaa agagttaata 3600acttctacag tcagggttgc
ccggctcctt tcctacctag taggctaacg atacacatgt 3660atctagggtg atgaactgac
ccgtgattga aaacggtaag gggtagagtc gagaggctct 3720accccatttc tttttctgga
ggaatgcaat tatgagcatc tacgcttttg atattgatat 3780taccggagta cttcataagc
aaggacctta tctacttggg gaaccagaaa cagtaagtaa 3840gttcattgct ccctggcaat
ctgtgttctt acctaactgt gaagagtccc atgagattgt 3900tggctgtctt ttcttatcaa
acctacaact caatactgag atacttaccg aaccaaaggt 3960gtatgacctc actcgttatg
acggattcaa tgagttcttc cctggtatac gtttctataa 4020aatacccggt tgggaattga
tagttgacca taacgtcttt aataaaacac tttccattta 4080cttcagacca ctttcctaaa
ggaaggagaa tcaatcatga aattcataat gaccatcccc 4140agttttgctg aaagggagtc
tatgtataca aatgacttct ctttagaaga agtcatgaat 4200agtagccccc aacatgtaat
cctagttcct accttagagg actaccatga agtaaggggc 4260cagttattct tagcaggtaa
tgaccgtaag gttgtatgta ttgtcccacc ttccgtctac 4320tcctttagtg atgcatatga
cgctctacct ttcatgatgg tatccgatga ataccaatgg 4380gatagtcgcc ataagaatga
agaataccat tactactatg taaggaagta agttatgcac 4440ttcttcggtt taagtagtcg
tagtacatgt ccccttaatc tgggagtagg taaatccccc 4500tggttgggaa taagtaaacc
catgaccatg tactatgaac cctgtaaaga agcagatgag 4560ttcataggaa cctgtctaat
gttaggtgca cctattacct ttattaagat aaatacttcc 4620atcgaagttc cagaggatgt
cgctgtatac atcttctcct ccagtgaacc tatggtagct 4680actactctta caggcagact
cattaccctc tatccttcct catatttatc tagcgacgga 4740agctactaat ggttatcagt
aaaatttatt cagagctacc tggggatgaa tacacccacc 4800ataaagaatt aactaacctg
gatgctgatg tactcatact ggttccagac gaacagtcct 4860tgcatgaact acaaggatta
ctcttcttat acaataagaa ttcggaagta catctacttg 4920aaccaggaac accgatgtcc
tggcagggta gcttcagtcc ttggcatctc aatacggaca 4980agtacatact atccttcgag
gataacaccc caatctataa ggaaagacaa tgaaactaaa 5040cacatcaccc tatccggtgc
agttagaagt agtactcgac cgggatacct tcattaaaaa 5100gtacaagaaa cttaaagggt
atgagccaga tttggaaggt tgtaaggggt atactacata 5160ctctgataat aaggttctta
tgggcatctt ctcagacccc ttgccaacgc ttatccatga 5220agtaaaccac ttctgcttat
gggtattcga ttatattggt atgcctataa acagtagtaa 5280tagtgaagcg tactgttact
acatggatag cattcttgaa caggtactga agaatgaacg 5340ttaaactatt aggtatgaaa
ggtactgact tactgcacta ttcctctgaa ccagggaaag 5400caatcattca cattatgtcc
tacggaacct caacgctaat aatggagcca ggagagttag 5460cagactttgc acagggacat
aagctaatta ctaacagtaa tgcagagata gttatcctgc 5520aaaatggtga gtcagtattc
ataccatacg gtgattatgt ctatgtattt agcaaagatg 5580ctaccgtaag aaatagtagt
ggtaatccta atcatccgga ctattactct ggtatagcac 5640gcatcatcac cagaaagccc
catcttacag gataacttac acaattaagc tcttaaggat 5700attaaatgaa acttactaag
ataggttatg ctgatggacc tatttccact acccatacaa 5760ccttacaaga ccttgtacct
ggtagactca ctcctattat cctagtacca gatgatcaga 5820ctctacacga agttaaagga
ttagctttac tactaaataa ggtgactatt gtagaagttc 5880ttgaaccagg aaaggaatta
ccgtggtcct cctacatgta ctccccctgg catgtgaata 5940cggataagta cactcttacc
tttggtatta ccagtgaact gcaaactcct cacttggtat 6000atgcttcctc ctatttaact
gcttataagt aggatagcta tcatattttt gcatatgaaa 6060agtagcatag ttatgtatgg
ctacagtggt gtggctgggg ctacaccctt acactcaacc 6120at
612233789DNAArtificial
Sequencea part of DNA of novel bacteriophage CJ23 Contig00003
3ccatgtggtt cgtactgtct actgtaactt aggtaatctt aaactcctgt atgatttctt
60aaatcagtac gggatggtta ttggtgtaca gtctgagata gaacttaaat cactccctct
120gaaaacaaag tatgcacgca tttatggcag gaaccgtagt aataaaagaa ccattaccga
180ctacttatat gttgatgatg ataaatctgg aatcataccg gatgacccgg cagctacagg
240ttcctgggta caagtatcca tgacctcaag tggtagtgag tccactaata caaataccta
300tatagttgtt gggtatacct ctgctggagg ggaaacttct ataccagttg agtttgatac
360cgttggcata ccttttatta cagtaggggg attcacacag ctcaatggga agggtttcac
420atattctact ggtgatactg agattaaatt agctcaggaa ctggaagaag gtgacgaagt
480tattatgttc cttacaggtg tgcctgcttc cccagatact gttgctattg ataactggaa
540ggttgttaac tggttataca attttggtaa tgctgtagga ggtgagcagg ttatagatat
600cccattcgct ttcattgatg tacctgctgt gtataagaat ggggcaagat tgtataaagg
660tttgcccaat aagtcataca ctgtcgatgc agagaataag agaatcttcc tgacagaacc
720acttgtaacc gatgataggg taattattac tattgggggc aatcaagaaa taatttatgt
780ttctgataga acaatccagg aagtagctcg cggattcaac ttacgggatt ctgagataat
840tttagatacg gatactgtta cctacttgaa tggtaaagtg gttgtgtatg tagcttctca
900gcagaagtca tacaagctcc ctacactacc aactaatgtc agaattaaat ctgtagtagg
960ggatttactc acttacgttc cgggtaacat tacagtttcc cttattccaa ttaacattat
1020ttaagcaaag ggtcactaag tgacctttaa tagaaaaaca aagaggttcg tatgaacgag
1080atgttcagtc aaggcggtaa aggttccact ggtattctta ccaacaagca agccattgcc
1140cgtaagtttg gtattaagca gaatgaagtg gtctactttt ctgtaggagt agacttgggt
1200ggatacaaag tcatttatga caagactact caacgtgctt actcattgcc tgtacttcca
1260gcaggaacca ttgctgtaag tctcagtgaa catgcagtct tggttcattc agcaggtaca
1320gttgatttgg gtgaactggc tgctgcacgt agagagtttg tatgcttatc tgattcattt
1380actacgggct tagtagtaaa tactcgaaat gagcttttga tgcataatgg tattggttat
1440acctacttag gttctctacc cgtaactatt gttgaaggga ccaaccctgt tggcaacacg
1500gactggaagt ctcaaacgga tcctaatttg cgtgctcagt tggcatctca ggatggtatg
1560accttaattg gtagtgtacc tgacgtaact gccctggctt ctgtcggagc aacagtaggg
1620tctagtgtaa tgctagattc atactcaggc tcttccgttg gtgggggtat tatgattgcg
1680gttcctaata ctactcctgt agatgaagtt gtaactttca ccggtgcagg tgtggtctgg
1740aagcgtaagt tctttgaagg tactgctacg gtatatgatg caggttatac tggtacgggg
1800gatattgccc cattcattaa caaggtaaac tctgccggtt acgattgtct tgtacctaca
1860tcagggacta tcagtacacc tatcttactg gatgtagcta agggggcttt agtaggagct
1920aataagtgta cccttacgga actggaaggt gtaacaggag agtattactt aaccataatc
1980aactctaata cggattacac agctcgtgat gccattaatg ctactgccct attaacaggt
2040atttcctttg taggtaaagg tactcgaaag atgtgtttgg gtagtagtac tggaggggag
2100atagcagaat tacgtatatc taactgtggg tttatatcta ccgcaggtat tgagtttaaa
2160gataatgcat accgtattct atttgataag tgcactattt ctcgcagttt tactaactct
2220gtaatcttta attccctggc taatgctggt gaggttataa aattcaacca ctgctggatg
2280gttgataatg ggggtccatt tacttttgag aatgggcagt tcatctttga ttcatgctca
2340ttaccagcag gtaaaaaggc aggctacttc gatccagtag tggcattaag tgataatgct
2400accctagtat ttgctaacgg taatattgag tatcaacctg ggcagagctt tgtaggcttt
2460actgtgagtg gaagctcacg cctcagtatt aaagattcta ctattctgct tccggaaggc
2520tacagtacag tgcctattgt tagtaatggt gacggggtag ttagtttaaa taactgctca
2580ttgcccctct acggtaacac aacgattgct actggatttg ccacaagaca gttgataggt
2640ggctctagca agaaagtaat gtctagaggg tgcttccctc gtgcaggctt tattacaact
2700aactggaatc taggaagcat tgtaagccct tatattaata gtattagcaa tggctcagga
2760cagtttgaaa atacatctaa ctggacactc tctcaaactg gaacaggtgc tgtcactgct
2820actacagcaa acgatgttcc taacgattta atgtttacaa cttcttttgt tttatctgta
2880ccatcagcag atgcagcagc taacttcact caaacaatca ttgattgtga accgggtcgt
2940tattttcagc ttggtttttg ggctaaaaat acaacaacca ccctggcgtc aattagattc
3000ctggaccagc aaggaaatgc tgttgcggat tccataggat atatcatccc agtggtaaac
3060acgttcaact tttacgcttt ggtggattgc gttcctccag gagcttacaa agctgaaatt
3120aattttaatg tttcctctgt tgttgggggc gtcgtaatac acaatgcagt ttacggattg
3180atttaattaa aactaaggcc caaactgggc cttttcttat atagagttta ctgtttgcat
3240taccctaacc gatactgctg ttagagtaat tacttgcccg gataagttaa acatacccaa
3300ctgaatacca tccaccatga atgggtcagt tgaaccaagt gtatatgtag ctaatacgtt
3360atctctgtta gcaatatccg taccttgtac tttaatacta gcgtctgaac ctactactgt
3420agttccattg gttcgtcttg tttgcgttct ccattccctt gctgtaccag caccacctgc
3480aatagttcct gagatacgaa cactaaacat tacctgggta ttcctggttc taacagggaa
3540cttaagatta ccttcttcaa ttgttaatgc tgctgtaccc ccaggtgatt tagttatcca
3600tggtaatgaa aagtaattga gccaggactg gtcatttact acctggctcc cggtccatgt
3660atacaagtcc agaaggaaag tatcttctaa ctctgttggt gggatgggta gtagttcagg
3720gggaagctct gaaacgttat tagcagaaac tattcgttcc ccatagcttc ccacaaccag
3780caaaccccc
3789
User Contributions:
Comment about this patent or add new information about this topic: